LOCUS       Exported                5610 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 5610)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
  JOURNAL   Exported from Poly.
REFERENCE   2  (bases 1 to 5610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 24, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5610
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..5610
                     /note="""transcription terminator for bacteriophage T7 RNA"
     CDS             complement(145..1227)
                     /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"
     promoter        complement(1228..1305)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        1618..1636
                     /label=T7 promoter
                     /note="t7 promoter"
     protein_bind    1637..1661
                     /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)."
     RBS             1676..1698
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1706..3424
                     /codon_start=1
                     /label=HIV-RT
                     /translation="MHHHHHHGGSGMPISPIETVPVKLKPGMDGPKVKQWPLTEEKIKA
                     LVEICTEMEKEGKISKIGPENPYNTPVFAIKKKDSTKWRKLVDFRELNKRTQDFWEVQL
                     GIPHPAGLKKKKSVTVLDVGDAYFSVPLDEDFRKYTAFTIPSINNETPGIRYQYNVLPQ
                     GWKGSPAIFQSSMTKILEPFRKQNPDIVIYQYMDDLYVGSDLEIGQHRTKIEELRQHLL
                     RWGLTTPDKKHQKEPPFLWMGYELHPDKWTVQPIVLPEKDSWTVNDIQKLVGKLNWASQ
                     IYPGIKVRQLCKLLRGTKALTEVIPLTEEAELELAENREILKEPVHGVYYDPSKDLIAE
                     IQKQGQGQWTYQIYQEPFKNLKTGKYARMRGAHTNDVKQLTEAVQKITTESIVIWGKTP
                     KFKLPIQKETWETWWTEYWQATWIPEWEFVNTPPLVKLWYQLEKEPIVGAETFYVDGAA
                     NRETKLGKAGYVTNRGRQKVVTLTDTTNQKTELQAIYLALQDSGLEVNIVTDSQYALGI
                     IQAQPDQSESELVNQIIEQLIKKEKVYLAWVPAHKGIGGNEQVDKLVSAGIRKVL"
     CDS             1709..1726
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3488..3535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3647..3694
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3699..3715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3782..4370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4561..5421)
                     /codon_start=1
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5422..5526)
                     /label=bla promoter
                     /note="ampicillin resistance gene promoter"
     primer_bind     5542..5558
                     /label=M13 for
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5559..5590
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ttaattaaca ttactcgcat ccattctcag gctgtctcgt ctcgtctccg gaaggagctg
       61 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
      121 cttacattaa ttgcgttgcg ctcatcactg cccgctttcc agtcgggaaa cctgtcgtgc
      181 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
      241 ggtggttttt cttttcacca gtgacacggg caacagctga ttgcccttca ccgcctggcc
      301 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
      361 gatggtggtt aacggcggga tataacatga gctatcttcg gtatcgtcgt atcccactac
      421 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
      481 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
      541 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
      601 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
      661 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
      721 cagtcgcgta ccatcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
      781 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
      841 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
      901 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
      961 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     1021 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     1081 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     1141 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     1201 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     1261 ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtccggga tctcgacgct
     1321 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     1381 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     1441 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     1501 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     1561 tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc gcgaaattaa
     1621 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     1681 ttaactttaa gaaggagata tacatatgca ccatcatcat catcatggtg gaagcggtat
     1741 gccgattagc ccgattgaaa ccgttccggt taaactgaaa cctggtatgg atggtccgaa
     1801 agttaaacag tggcctctga ccgaagaaaa aatcaaagca ctggtggaaa tttgcaccga
     1861 gatggaaaaa gaaggcaaaa tcagcaaaat tggtccggaa aatccgtata atacaccggt
     1921 ttttgccatc aagaaaaagg atagcaccaa atggcgtaaa ctggtggatt ttcgtgaact
     1981 gaataaacgc acccaggatt tttgggaagt tcagctgggt attccgcatc cggcaggtct
     2041 gaaaaaaaag aaaagcgtta cagttctgga tgtgggtgat gcatatttta gcgttccgct
     2101 ggatgaagat tttcgtaaat ataccgcatt tacgatcccg agcattaata acgaaacacc
     2161 gggtattcgc tatcagtata atgttctgcc gcaaggttgg aaaggtagtc cggcaatttt
     2221 tcagagcagc atgaccaaaa ttctggaacc gtttcgtaaa cagaatccgg atattgtgat
     2281 ctaccagtat atggatgatc tgtatgttgg tagcgatctg gaaattggtc agcatcgtac
     2341 aaaaattgaa gaactgcgtc agcatctgct gcgttggggt ctgaccacac cggataaaaa
     2401 acatcagaaa gaaccgcctt ttctgtggat gggttatgaa ctgcatccgg ataaatggac
     2461 cgttcagccg attgtgctgc cggaaaaaga tagctggacc gttaatgata ttcagaaact
     2521 ggttggcaaa ctgaattggg caagccagat ctatccgggt attaaagttc gtcagctgtg
     2581 taaactgctg cgtggtacaa aagcactgac agaagttatt ccgctgacgg aagaagcaga
     2641 actggaactg gcagaaaatc gtgaaattct gaaagaacct gtgcacggcg tttattatga
     2701 tccgagcaaa gatctgattg ccgagatcca gaaacaaggt caaggtcagt ggacctatca
     2761 gatttatcaa gaaccgttca agaatctgaa aaccggtaaa tatgcacgta tgcgtggtgc
     2821 acataccaat gatgtgaaac agctgactga agccgttcag aaaatcacaa ccgaaagcat
     2881 tgttatttgg ggtaaaaccc cgaaattcaa actgccgatt cagaaagaga catgggaaac
     2941 ttggtggacc gaatattggc aggcaacctg gattcctgaa tgggaatttg ttaatacccc
     3001 tccgctggtg aaactgtggt atcagttaga aaaagaaccg attgttggtg ccgaaacctt
     3061 ttatgttgat ggtgcagcaa atcgcgaaac caaactgggt aaagcaggtt atgttaccaa
     3121 ccgtggccgc cagaaggtag ttactttaac ggacaccacg aatcagaaaa cagaattaca
     3181 agcgatttac ttggcgcttc aagatagcgg tctggaagta aacatcgtta cggacagcca
     3241 gtacgcactt gggattattc aggcccagcc ggatcagtct gaaagtgaac tggtaaatca
     3301 aattattgag cagctgatta aaaaggagaa ggtgtatctg gcttgggtcc cggctcataa
     3361 aggtattgga gggaacgagc aagttgacaa actggtttct gcgggtattc gcaaggtcct
     3421 ttaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3481 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     3541 aggaggaact atatccggat tccccaggat acatagatta ccacaactcc gagcccttcc
     3601 accactagtc gctgaggtgt caatcgtcgg agccgctgag caataactag cataacccct
     3661 tggggcctct aaacgggtct tgaggggttt tttgcatggt catagctgtt tcctgagagc
     3721 ttggcaggtg atgacacaca ttaacaaatt tcgtgaggag tctccagaag aatgccatta
     3781 atttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3961 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4201 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaaggcctac
     4381 tattagcaac aacgatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcacc tgcaccagtc agtaaaacga cggccagtag tcaaaagcct ccgaccggag
     5581 gcttttgact tggttcaggt ggagtgggag
//
