LOCUS       sequence-459281-        4744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4744)
  AUTHORS   Guet CC, Bruneaux L, Min TL, Siegal-Gaskins D, Figueroa I, Emonet T,
            Cluzel P
  TITLE     Minimally invasive determination of mRNA concentration in single 
            living bacteria.
  JOURNAL   Nucleic Acids Res. 2008 Jul . 36(12):e73.
  PUBMED    18515347
REFERENCE   2  (bases 1 to 4744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2008 Jul . 36(12):e73."
COMMENT     Sequence Label: pZS*12-MS2-GFP
FEATURES             Location/Qualifiers
     source          1..4744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             375..1088
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(405..433)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1028..1048
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1118..1204
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1742..2692)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2740..2962
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3454..3548)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3578..4369,4370..4438))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4201..4220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4439..4543)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4611..4629
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    4652..4668
                     /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)."
     protein_bind    4675..4691
                     /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             4733..4744
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ggtaccgcat ggcttctaac tttactcagt tcgttctcgt cgacaatggc ggaactggcg
       61 acgtgactgt cgccccaagc aacttcgcta acggggtcgc tgaatggatc agctctaact
      121 cgcgttcaca ggcttacaaa gtaacctgta gcgttcgtca gagctctgcg cagaatcgca
      181 aatacaccat caaagtcgag gtgcctaaag gcgcatggcg ttcgtactta aatatggaac
      241 taaccattcc aattttctcc acgaattccg actgcgagct tattgttaag gcaatgcaag
      301 gtctcctaaa agatggaaac ccgattccct cagcaatcgc agcaaactcc ggcatctacg
      361 gcggtggagg ttccatgcgt aaaggagaag aacttttcac tggagttgtc ccaattcttg
      421 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
      481 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
      541 catggccaac acttgtcact actttcggtt atggtgttca atgctttgcg agatacccag
      601 atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa
      661 gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag
      721 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
      781 ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca
      841 aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg
      901 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
      961 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     1021 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     1081 tatacaaata agcttaatta gctgagtcta gaggcatcaa ataaaacgaa aggctcagtc
     1141 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     1201 aaatccgccg ccctagacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt
     1261 gttgctagtt tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg
     1321 ctatttcttc cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt
     1381 tgtcggcagc tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg
     1441 ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact
     1501 ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc
     1561 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
     1621 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
     1681 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
     1741 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
     1801 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
     1861 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
     1921 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
     1981 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
     2041 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
     2101 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
     2161 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     2221 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     2281 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     2341 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     2401 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     2461 tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca
     2521 tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa
     2581 tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt
     2641 catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca
     2701 attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac
     2761 tagtcctttt cccgggagat ctgggtatct gtaaattctg ctagaccttt gctggaaaac
     2821 ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt
     2881 atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag
     2941 atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt
     3001 cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca
     3061 ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg
     3121 agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg
     3181 ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa
     3241 gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc
     3301 tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt
     3361 atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac
     3421 aggcttaccc gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg
     3481 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     3541 caggagtcca agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4561 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4621 tcacgaggcc ctttcgtctt cacctcgaga attgtgagcg gataacaatt gacattgtga
     4681 gcggataaca agatactgag cacatcagca ggacgcactg accgaattca ttaaagagga
     4741 gaaa
//
