LOCUS       Exported                6930 bp ds-DNA     circular SYN 24-SEP-2020
DEFINITION  synthetic linear DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6930)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
  JOURNAL   Exported from Poly.
REFERENCE   2  (bases 1 to 6930)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 24, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6930
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..6930
                     /label=T7 terminator
                     /note="""transcription terminator for bacteriophage T7 RNA"
     CDS             716..907
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1009..1151
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1337..1925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2047..2862
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(2955..3410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     terminator      complement(3447..3494)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3558..5276)
                     /codon_start=1
                     /label=HIV-RT
                     /translation="MHHHHHHGGSGMPISPIETVPVKLKPGMDGPKVKQWPLTEEKIKA
                     LVEICTEMEKEGKISKIGPENPYNTPVFAIKKKDSTKWRKLVDFRELNKRTQDFWEVQL
                     GIPHPAGLKKKKSVTVLDVGDAYFSVPLDEDFRKYTAFTIPSINNETPGIRYQYNVLPQ
                     GWKGSPAIFQSSMTKILEPFRKQNPDIVIYQYMDDLYVGSDLEIGQHRTKIEELRQHLL
                     RWGLTTPDKKHQKEPPFLWMGYELHPDKWTVQPIVLPEKDSWTVNDIQKLVGKLNWASQ
                     IYPGIKVRQLCKLLRGTKALTEVIPLTEEAELELAENREILKEPVHGVYYDPSKDLIAE
                     IQKQGQGQWTYQIYQEPFKNLKTGKYARMRGAHTNDVKQLTEAVQKITTESIVIWGKTP
                     KFKLPIQKETWETWWTEYWQATWIPEWEFVNTPPLVKLWYQLEKEPIVGAETFYVDGAA
                     NRETKLGKAGYVTNRGRQKVVTLTDTTNQKTELQAIYLALQDSGLEVNIVTDSQYALGI
                     IQAQPDQSESELVNQIIEQLIKKEKVYLAWVPAHKGIGGNEQVDKLVSAGIRKVL"
     CDS             complement(5256..5273)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5284..5306
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5321..5345
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5346..5364)
                     /label=T7 promoter
                     /note="t7 promoter"
     promoter        5677..5754
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5755..6837
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
       61 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
      121 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
      181 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
      241 ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
      301 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
      361 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
      421 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
      481 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
      541 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
      601 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
      661 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
      721 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
      781 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
      841 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
      901 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      961 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     1021 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     1081 gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1141 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1741 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1981 gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2041 ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac
     2101 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2161 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2221 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2281 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2341 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2401 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2461 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2521 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2581 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2641 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2701 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2761 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2821 aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg
     2881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2941 aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     3001 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3061 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3121 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3181 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3241 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3301 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3361 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     3421 aatccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca
     3481 aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct
     3541 ttgttagcag ccggatctta aaggaccttg cgaatacccg cagaaaccag tttgtcaact
     3601 tgctcgttcc ctccaatacc tttatgagcc gggacccaag ccagatacac cttctccttt
     3661 ttaatcagct gctcaataat ttgatttacc agttcacttt cagactgatc cggctgggcc
     3721 tgaataatcc caagtgcgta ctggctgtcc gtaacgatgt ttacttccag accgctatct
     3781 tgaagcgcca agtaaatcgc ttgtaattct gttttctgat tcgtggtgtc cgttaaagta
     3841 actaccttct ggcggccacg gttggtaaca taacctgctt tacccagttt ggtttcgcga
     3901 tttgctgcac catcaacata aaaggtttcg gcaccaacaa tcggttcttt ttctaactga
     3961 taccacagtt tcaccagcgg aggggtatta acaaattccc attcaggaat ccaggttgcc
     4021 tgccaatatt cggtccacca agtttcccat gtctctttct gaatcggcag tttgaatttc
     4081 ggggttttac cccaaataac aatgctttcg gttgtgattt tctgaacggc ttcagtcagc
     4141 tgtttcacat cattggtatg tgcaccacgc atacgtgcat atttaccggt tttcagattc
     4201 ttgaacggtt cttgataaat ctgataggtc cactgacctt gaccttgttt ctggatctcg
     4261 gcaatcagat ctttgctcgg atcataataa acgccgtgca caggttcttt cagaatttca
     4321 cgattttctg ccagttccag ttctgcttct tccgtcagcg gaataacttc tgtcagtgct
     4381 tttgtaccac gcagcagttt acacagctga cgaactttaa tacccggata gatctggctt
     4441 gcccaattca gtttgccaac cagtttctga atatcattaa cggtccagct atctttttcc
     4501 ggcagcacaa tcggctgaac ggtccattta tccggatgca gttcataacc catccacaga
     4561 aaaggcggtt ctttctgatg ttttttatcc ggtgtggtca gaccccaacg cagcagatgc
     4621 tgacgcagtt cttcaatttt tgtacgatgc tgaccaattt ccagatcgct accaacatac
     4681 agatcatcca tatactggta gatcacaata tccggattct gtttacgaaa cggttccaga
     4741 attttggtca tgctgctctg aaaaattgcc ggactacctt tccaaccttg cggcagaaca
     4801 ttatactgat agcgaatacc cggtgtttcg ttattaatgc tcgggatcgt aaatgcggta
     4861 tatttacgaa aatcttcatc cagcggaacg ctaaaatatg catcacccac atccagaact
     4921 gtaacgcttt tctttttttt cagacctgcc ggatgcggaa tacccagctg aacttcccaa
     4981 aaatcctggg tgcgtttatt cagttcacga aaatccacca gtttacgcca tttggtgcta
     5041 tcctttttct tgatggcaaa aaccggtgta ttatacggat tttccggacc aattttgctg
     5101 attttgcctt ctttttccat ctcggtgcaa atttccacca gtgctttgat tttttcttcg
     5161 gtcagaggcc actgtttaac tttcggacca tccataccag gtttcagttt aaccggaacg
     5221 gtttcaatcg ggctaatcgg cataccgctt ccaccatgat gatgatgatg gtgcatatgt
     5281 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
     5341 attcccctat agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg
     5401 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     5461 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     5521 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     5581 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     5641 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     5701 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     5761 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     5821 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     5881 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     5941 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     6001 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     6061 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     6121 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     6181 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     6241 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     6301 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     6361 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     6421 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     6481 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     6541 cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     6601 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     6661 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     6721 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6781 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6841 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     6901 gagagccttc aacccagtca gctccttccg
//
