LOCUS       sequence-113709-        4357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express yellow fluorescent protein (YFP) with strong T7 RBS at a low
            gene dosage (3-4 copies/cell).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4357)
  AUTHORS   Subramaniam AR, Pan T, Cluzel P
  TITLE     Environmental perturbations lift the degeneracy of the genetic code 
            to regulate protein levels in bacteria.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Feb 5;110(6):2419-24. doi: 
            10.1073/pnas.1211077110. Epub 2012 Dec 31.
  PUBMED    23277573
REFERENCE   2  (bases 1 to 4357)
  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"; doi: 
            "10.1073/pnas.1211077110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-02-5- 5"; volume: "110"; issue: "6"; pages: "2419-24"
COMMENT     Sequence Label: pZS*11-yfp0
FEATURES             Location/Qualifiers
     source          1..4357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(6..24,25..30,31..47,48..53,54..79)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    6..24
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             92..114
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      867..953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1492..2442)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2490..2712
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3204..3298)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3328..4119,4120..4188))
                     /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     3951..3970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4189..4293)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg aattcaataa ttttgtttaa ctttaagaag gagatggtac
      121 catgagcaaa ggtgaagaac tgttcaccgg cgttgtgcca attctggttg agctggatgg
      181 tgacgtgaat ggccacaaat tttccgtgtc tggtgaaggc gagggtgatg ctacttatgg
      241 caaactgact ctgaaactga tctgtaccac cggcaaactg cctgttccat ggccaactct
      301 ggtcactact ctgggttacg gcctgatgtg ttttgcgcgt tacccggatc acatgaaaca
      361 gcatgacttc ttcaaatctg ccatgccgga aggctatgtc caagaacgta cgatcttttt
      421 caaggacgac ggcaactata aaacccgtgc cgaagttaaa ttcgagggtg acaccctggt
      481 taaccgcatc gaactgaaag gcattgactt caaagaggac ggcaacattc tgggtcacaa
      541 gctggaatac aactacaact cccacaacgt ttacattact gctgacaagc agaaaaacgg
      601 catcaaagca aacttcaaga tccgtcacaa cattgaagat ggtggcgtac agctggcaga
      661 tcactaccag cagaacactc caatcggtga tggcccagta ctgctgccag ataaccatta
      721 cctgtcctac cagagcaaac tgtctaaaga cccgaacgaa aaacgtgacc acatggtact
      781 gctggaattt gttaccgcgg caggcattac ccacggtatg gacgaactgt ataaatagaa
      841 gcttaattag ctgagtctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc
      961 cctagaccta gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt
     1021 gttatcagaa tcgcagatcc ggcttcagcc ggttttgccg gctgaaagcg ctatttcttc
     1081 cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc
     1141 tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta
     1201 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     1261 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     1321 gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     1381 tcatctgtgc atatggacag ttttcccttt gatatctaac ggtgaacagt tgttctactt
     1441 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     1501 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     1561 cgaaccattg agatcatgct tactttgcat gtcactcaaa aattttgcct caaaactggt
     1621 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta
     1681 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     1741 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     1801 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     1861 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     1921 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     1981 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     2041 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     2101 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     2161 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     2221 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     2281 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     2341 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     2401 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     2461 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     2521 cccgggagat ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     2581 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     2641 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     2701 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     2761 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     2821 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     2881 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     2941 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     3001 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     3061 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     3121 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc
     3181 gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     3241 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     3301 agcgagctcg taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4321 cgtccaacat atggcaccgg aggctttcgt cttcacc
//
