LOCUS       Exported                4258 bp DNA     circular SYN 04-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4258)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 4, 2023 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..109
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    110..133
                     /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)."
     misc_feature    131..166
                     /label=target seq
     CDS             180..893
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             950..994
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      1046..1093
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1264..2055)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2056..2147)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2195..2933)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3143..4225)
                     /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(join(4226..4258,1..45))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
       61 ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag
      121 cggataacaa ttcgccatgc tcaggctggc gagtgcgcca ctcatcaagg agatatacca
      181 tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg
      241 atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta
      301 aactgacgct gaagttcatc tgtactactg gtaaactgcc ggttccttgg ccgactctgg
      361 taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc
      421 atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta
      481 aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa
      541 accgcattga gctgaaaggc attgacttta aagaggacgg caatatcctg ggccataagc
      601 tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca
      661 ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc
      721 actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc
      781 tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc
      841 tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgaggat
      901 ccaagccagt gataagtgga atgccatgtg ggctgtcctc gagtctggta aagaaaccgc
      961 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     1021 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1081 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     1141 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     1201 accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac gaattgttag
     1261 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     1321 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     1381 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     1441 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     1501 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     1561 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     1621 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     1681 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     1741 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     1801 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     1861 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     1921 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     1981 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     2041 caccgcttcc ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag ctagctcact
     2161 cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata caaagttacc
     2221 cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag ggccgcgccg
     2281 gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag acgcttttcc
     2341 ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg cgaaacccga
     2401 caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc
     2461 gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag tgtggcgctt
     2521 tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc aagctgggct
     2581 gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac tgttcacttg
     2641 agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt aactgggagt
     2701 tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg
     2761 ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac cacggttaag
     2821 cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt ttttcgttta
     2881 cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2941 ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     3001 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     3061 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3121 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3241 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3301 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3361 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3421 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3481 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3541 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3601 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3661 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3721 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3781 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     3841 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     3901 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     3961 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4021 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4081 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4141 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4201 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcg
//
