LOCUS       Exported                4404 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 4404)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
  JOURNAL   Exported from Poly.
REFERENCE   2  (bases 1 to 4404)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Sep 24, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4404
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..4404
                     /label=T7 terminator
                     /note="""transcription terminator for bacteriophage T7 RNA"
     CDS             complement(118..1200)
                     /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(1201..1278)
                     /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        1587..1605
                     /label=T7 promoter
                     /note="t7 promoter"
     protein_bind    1606..1630
                     /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             1645..1667
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1674..2264
                     /codon_start=1
                     /label=EcPPA Pyrophosphatase
                     /translation="MGSSHHHHHHSSGLVPRGSHMSLLNVPAGKDLPEDIYVVIEIPAN
                     ADPIKYEIDKESGALFVDRFMSTAMFYPCNYGYINHTLSLDGDPVDVLVPTPYPLQPGS
                     VIRCRPVGVLKMTDEAGEDAKLVAVPHSKLSKEYDHIKDVNDLPELLKAQIAHFFEHYK
                     DLEKGKWVKVEGWENAEAAKAEIVASFERAKNK"
     CDS             1686..1703
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1713..1730
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2331..2378
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2441..2488
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2493..2509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2576..3164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3355..4215)
                     /codon_start=1
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4216..4320)
                     /label=bla promoter
                     /note="ampicillin resistance gene promoter"
     primer_bind     4336..4352
                     /label=M13 for
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4353..4384
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
       61 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
      121 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      181 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgacac
      241 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
      301 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
      361 tgagctatct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
      421 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
      481 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
      541 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
      601 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
      661 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccatctt catgggagaa
      721 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
      781 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
      841 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
      901 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
      961 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     1021 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     1081 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     1141 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     1201 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     1261 gcaccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     1321 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     1381 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     1441 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     1501 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     1561 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     1621 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
     1681 gcagccatca tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatgagct
     1741 tactcaacgt ccctgcgggt aaagatctgc cggaagatat ctacgttgtt attgagatcc
     1801 cggctaacgc agatccgatc aaatacgaaa tcgacaaaga gagcggcgca ctgttcgttg
     1861 accgcttcat gtccaccgcg atgttctatc catgcaacta cggttacatc aaccacaccc
     1921 tgtctctgga cggtgacccg gttgacgtac tggtcccaac tccgtacccg ctgcagccgg
     1981 gttctgtgat ccgttgccgt ccggttggcg ttctgaaaat gaccgacgaa gccggtgaag
     2041 atgcgaaact ggttgcggtt ccgcacagca agctgagcaa agaatacgat cacattaaag
     2101 acgttaacga tctgcctgaa ctgctgaaag cgcaaatcgc tcacttcttc gagcactaca
     2161 aagacctcga aaaaggcaag tgggtgaaag ttgaaggttg ggaaaacgca gaagccgcta
     2221 aagctgaaat cgttgcctcc ttcgagcgcg caaagaataa ataatgagat ccggctgcta
     2281 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     2341 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     2401 gatcgctgag gtgtcaatcg tcggagccgc tgagcaataa ctagcataac cccttggggc
     2461 ctctaaacgg gtcttgaggg gttttttgca tggtcatagc tgtttcctga gagcttggca
     2521 ggtgatgaca cacattaaca aatttcgtga ggagtctcca gaagaatgcc attaatttcc
     2581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaaggc ctactattag
     3181 caacaacgat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3481 tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4321 cacctgcacc agtcagtaaa acgacggcca gtagtcaaaa gcctccgacc ggaggctttt
     4381 gacttggttc aggtggagtg ggag
//
