LOCUS       sequence-311103-        5442 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses wild-type gas vesicle protein C (GvpC) from Anabaena 
            flos-aquae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  AUTHORS   Lakshmanan A, Jin Z, Nety SP, Sawyer DP, Lee-Gosselin A, Malounda D,
            Swift MB, Maresca D, Shapiro MG
  TITLE     Acoustic biosensors for ultrasound imaging of enzyme activity.
  JOURNAL   Nat Chem Biol. 2020 Sep;16(9):988-996. doi: 
            10.1038/s41589-020-0591-0. Epub 2020 Jul 13.
  PUBMED    32661379
REFERENCE   2  (bases 1 to 5442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-020-0591-0"; journalName: "Nat Chem Biol"; date: 
            "2020-09"; volume: "16"; issue: "9"; pages: "988-996"
COMMENT     Sequence Label: pTetR-BCD2-Ana GvpC
FEATURES             Location/Qualifiers
     source          1..5442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(37..663)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     568..587
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        679..734
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    715..733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    775..793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    800..818
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             919..927
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      1544..1571
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1658..1675)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1828..1914
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2006..2033
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2052..2143
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2601..3056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2688..2707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2898..2919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3612..4271)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4186..4205
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4272..4374)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4766..4783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4900..5442,1..3))
                     /direction=LEFT
                     /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.
                     "
     primer_bind     complement(4980..4999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caagaagatc atcttattaa tcagataaaa tatttgttaa gacccacttt cacatttaag
       61 ttgtttttct aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcacc
      121 ttggtgatca aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg
      181 tgtttccctt tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt
      241 aaaatgcccc acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca
      301 taaaaaggct aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa
      361 atgtactttt gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt
      421 acgtaaaaaa tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc
      481 taacatctca atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa
      541 tgtaggctgc tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc
      601 gacctcatta agcagctcta atgcgctgtt aatcacttta cttttatcta atcttgacat
      661 cattaattcc taatttttgt tgacactcta tcgttgatag agttatttta ccactcccta
      721 tcagtgatag agaaaagtga aaatccatat gactagcaga tccactagag tcgatcccta
      781 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacct ctctactgtt
      841 tctccatagg gcccaagttc acttaaaaag gagatcaaca atgaaagcaa ttttcgtact
      901 gaaacatctt aatcatgcta aggaggtttt ctaatgattt ctttaatggc aaaaatccgg
      961 caagaacatc agtcaatagc agagaaagtg gctgaactat ctcttgagac cagagaattc
     1021 ttgtccgtca cgacagcgaa aagacaagag caagctgaaa aacaagctca agaactgcaa
     1081 gcattctaca aggatcttca ggaaacaagt cagcagtttt tatcagaaac agcccaagcc
     1141 agaattgctc aagctgaaaa acaagctcaa gaactgttag cattccacaa agaacttcaa
     1201 gaaacaagtc agcagttttt atcagcaaca gcccaagcca gaattgctca agctgaaaaa
     1261 caagcgcaag aactgttagc attttatcaa gaagttcggg aaacaagtca gcagttttta
     1321 tcagcaacag cccaagcaag aattgctcaa gctgaaaaac aagctcaaga actgttagca
     1381 ttccacaaag aacttcaaga aacaagtcag cagtttttat cagcaacagc cgacgcaaga
     1441 actgctcaag ctaaggaaca gaaggaatct ctcctgaaat tccgtcagga tttgtttgtg
     1501 agtatctttg gttaataata ctagatgcat taataatcac actggctcac cttcgggtgg
     1561 gcctttctgc gtttatatac tagagagaga atataaaaag ccagattatt aatccggctt
     1621 ttttattatt tcctgccgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     1681 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     1741 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1801 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1861 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1921 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     1981 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
     2041 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2101 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2161 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2221 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcagcaaact attaactggc
     2281 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2341 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2401 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2461 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2521 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2581 tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2941 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3001 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaaa
     3061 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3121 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3181 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3241 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3301 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3361 caccgcctac atacctcgct ctgctaatcc tgttaccagt caggcatttg agaagcacac
     3421 ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat
     3481 ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc
     3541 atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca ccaataactg
     3601 ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     3661 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     3721 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     3781 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     3841 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     3901 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     3961 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     4021 atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg
     4081 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     4141 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     4201 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     4261 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     4321 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct
     4381 cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt
     4441 tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat
     4501 gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt
     4561 ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac
     4621 cgccggacat cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt
     4681 gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata
     4741 tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     4801 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     4861 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     4921 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
     5041 gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca
     5101 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
     5161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
     5221 caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
     5281 ggttaaggct aaactgaaag gacaagtttt ggtgactacg ctcctccaag ccagttacct
     5341 cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt
     5401 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ct
//
