LOCUS       Exported                5832 bp ds-DNA     circular SYN 30-APR-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 30, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5832
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..29
                     /label=rrnC term
                     /note="color: #ff58aa"
     gene            complement(39..113)
                     /label=Ma Pyl tRNA(6)
                     /label=Mb PylTwt (NC_007355.1)
                     /note="color: #15af04; direction: LEFT"
     promoter        128..152
                     /label=Lpp promoter
                     /label=Ipp p
                     /note="color: #ff58aa"
     terminator      548..575
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      667..753
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     CDS             complement(992..1009)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             complement(1016..1729)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 2 segments:
                        1: 1016 .. 1726 / #05fd14
                        2: 1727 .. 1729 / #05fd14 / 1a"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(1745..2041)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="color: #cc99b2
                     Cleavage site after base 1744"
                     /translation="MGSDSEVNQEAKPEVKPEVKPETHINLKVSDGSS*IFFKIKKTTP
                     LRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     promoter        complement(2075..2359)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
                     /note="color: #ffffff; direction: LEFT"
     CDS             2386..3264
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /note="color: #993366"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      3552..4097
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
                     /note="color: #ffff00"
     CDS             complement(4393..5583)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /note="color: #ccffcc"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(5631..5659)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
                     /note="This reverse directional feature has 3 segments:
                        1: 5631 .. 5636 / #ffffff / -10
                        2: 5637 .. 5653 / #ffffff
                        3: 5654 .. 5659 / #ffffff / -35"
     primer_bind     5807..5823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
ORIGIN
        1 aaaaaaaatc cttagctttc gctaaggata ctagttggcg agagaccggg gtgtcgaacc
       61 ccgctatgct aggttttaga gacccgctgg tcgccggacc gtcccccaga tctagcgtta
      121 caagtattac acaaagtttt ttatgttgag aatatttttt tgatggggcg ccacttattt
      181 ttgatcgttc gctcaaagaa gcggcgccag ggttgttttt cttttcacca gtgagacggg
      241 caacagaacg ccatgagcgg cctcatttct tattctgagt tacaacagtc cgcaccgctg
      301 ccggtagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
      361 tctttatcat gcaactcgta ggacaggtgc cggcagcgcc caacagtccc ccggccacgg
      421 ggcctgccac catacccacg ccgaaacaag cgccctgcac cattatgttc cggatctgca
      481 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtgtcgaca gagtttgtag
      541 aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
      601 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
      661 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      721 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
      781 ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt
      841 caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc
      901 gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagccaagct
      961 tcgaattccc atatggtacc agctgcagtt aatggtgatg atgatggtgg ctgcctttat
     1021 acagttcatc cataccgtgg gtaatgcccg cggcggtaac aaattccagc agcaccatat
     1081 gatcacgttt ttcattcgga tctttgctca gaacgctctg ggtgctcaga taatgattat
     1141 ccggcagcag caccgggcca tcaccaatcg gggtattctg ctgataatga tccgccagct
     1201 gcacgctacc atcttccacg ttgtgacgga ttttaaagtt cgctttgatg ccatttttct
     1261 gtttatcggc ggtaatatac acattatggc tgttgaaatt atattccagt ttatgaccca
     1321 gaatgttgcc atcttcttta aaatcaatac ctttcagttc aatgcggttc accagggtat
     1381 cgccttcaaa tttaacttcc gcacgggttt tataggtgcc atcatctttg aagctaatgg
     1441 tacgttcctg cacatagcct tccggcatcg cgcttttaaa gaaatcatgg cgtttcatat
     1501 gatccggata gcggctaaag cactgaacgc cataggtcag ggtggtcacc agggtcggcc
     1561 acggaaccgg cagtttaccg gtggtgcaaa taaatttcag ggtcagttta ccgttggtcg
     1621 catcaccttc gccttcgcca cgaacgctaa atttatggcc attcacatca ccatccagtt
     1681 ccaccagaat cggcacaacg ccggtaaaca gttcttcacc tttgctaaca gagccggtac
     1741 cggatccacc aatctgttct ctgtgagcct caataatatc gttatcctcc atgtccaaat
     1801 cttcaggggt ctgatcagct tggattctaa taccgtcgta caagaatctt aaggagtcca
     1861 tttccttacc ctgtctttta gcgaacgctt ccatcagcct tcttaaagga gtggtctttt
     1921 tgatcttgaa gaagatctat gaagatccat cggacacctt taaattgatg tgagtctcag
     1981 gcttgacttc tggcttgacc tctggcttag cttcttgatt gacttctgag tcgctgccca
     2041 tggttaattc ctcctgttag cccaaaaaac gggtatggag aaacagtaga gagttgcgat
     2101 aaaaagcgtc aggtaggatc cgctaatctt atggataaaa atgctatggc atagcaaagt
     2161 gtgacgccgt gcaaataatc aatgtggact tttctgccgt gattatagac acttttgtta
     2221 cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa tgcttttaat aagcggggtt
     2281 accggtttgg ttagcgagaa gagccagtaa aagacgcagt gacggcaatg tctgatgcaa
     2341 tatggacaat tggtttcttc tctgaatggc gggagtatga aaagtatggc tgaagcgcaa
     2401 aatgatcccc tgctgccggg atactcgttt aatgcccatc tggtggcggg tttaacgccg
     2461 attgaggcca acggttatct cgattttttt atcgaccgac cgctgggaat gaaaggttat
     2521 attctcaatc tcaccattcg cggtcagggg gtggtgaaaa atcagggacg agaatttgtt
     2581 tgccgaccgg gtgatatttt gctgttcccg ccaggagaga ttcatcacta cggtcgtcat
     2641 ccggaggctc gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc ctactggcat
     2701 gaatggctta actggccgtc aatatttgcc aatacggggt tctttcgccc ggatgaagcg
     2761 caccagccgc atttcagcga cctgtttggg caaatcatta acgccgggca aggggaaggg
     2821 cgctattcgg agctgctggc gataaatctg cttgagcaat tgttactgcg gcgcatggaa
     2881 gcgattaacg agtcgctcca tccaccgatg gataatcggg tacgcgaggc ttgtcagtac
     2941 atcagcgatc acctggcaga cagcaatttt gatatcgcca gcgtcgcaca gcatgtttgc
     3001 ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt tagggattag cgtcttaagc
     3061 tggcgcgagg accaacgtat cagccaggcg aagctgcttt tgagcaccac ccggatgcct
     3121 atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac tctatttctc gcgggtattt
     3181 aaaaaatgca ccggggccag cccgagcgag ttccgtgccg gttgtgaaga aaaagtgaat
     3241 gatgtagccg tcaagttgtc ataattggta acgaatcaga caattgacgg cttgacggag
     3301 tagcataggg tttgcagaat ccctgcttcg tccatttgac aggcacatta tgcatgccgc
     3361 ttcgccttcg cgcgcgaatt gatctgctgc ctcgcgcgtt tcggtgatga cggtgagatc
     3421 ctgggcgcgg ggcatgacta acatgagaat tacaacttat atcgtatggg gctgacttca
     3481 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     3541 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     3601 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     3661 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     3721 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     3781 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3841 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3901 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3961 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     4021 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     4081 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     4141 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     4201 aacgaccgag cgtagcgagt cagtgagcga ggaagcggcc cgagatgcgc cgcgtgcggc
     4261 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag
     4321 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc
     4381 gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag
     4441 gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga
     4501 ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc
     4561 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc
     4621 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat
     4681 ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc
     4741 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     4801 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     4861 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     4921 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     4981 gctgactggg ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg
     5041 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     5101 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     5161 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     5221 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5281 ggcgtagagg atccacagga cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc
     5341 aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac
     5401 gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc
     5461 gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
     5521 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
     5581 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
     5641 atcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg ctgacttcag
     5701 gtgctacatt tgcaggagtc gcataaggga gagcgtctgg cgaaaggggg atgtgctgca
     5761 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     5821 agtgccaagc tt
//
