LOCUS       sequence-13254-e        5915 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 5915)
  AUTHORS   Tan C, Marguet P, You L
  TITLE     Emergent bistability by a growth-modulating positive feedback 
            circuit.
  JOURNAL   Nat Chem Biol. 2009 Nov . 5(11):842-8.
  PUBMED    19801994
REFERENCE   2  (bases 1 to 5915)
  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: "Nat Chem 
            Biol. 2009 Nov . 5(11):842-8."
COMMENT     Sequence Label: p15aTetT7mut-fuseYFP
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..96
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        join(248..266,267..272,273..289,290..295,296..321)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    248..266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    273..291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             331..342
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3014..3016,3017..3730)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3056..3077)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3317..3336)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3664..3685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3778..3864
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3900..3917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4579)
                     /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(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4693..4787)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4818..5612)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4929..4948)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5539..5558
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    5764..5780
                     /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)."
     primer_bind     5769..5791
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 gaaaaggccc ccaaggtagt tatccttaaa aaagccacag catacatcct gtccgtccaa
       61 gcagaggagc aaaagctcat ttctgaagag gacttgttgc ggaaacgacg agaacagttg
      121 aaacacaaac ttgaacagct acggaactct tgtgcgtaag gaaaagtaag gaaaacgatt
      181 ccttctaaca gaaatgtcct gagcaatcac ctatgaagct agcaggccct ttcgtcttca
      241 cctcgagtcc ctatcagtga tagagattga catccctatc agtgatagag atactgagca
      301 catcagcagg acgcactgac cgaattcatt aaagaggaga aagctaccgc atgaacacga
      361 ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg ttcaacactc
      421 tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag catgagtctt
      481 acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa gctgtgaggt
      541 tgcggataac gctgccgcca agcctctcat cactacccta ctccctaaga tgattgcacg
      601 catcaacgac tggtttgagg aagtgaaagc taagcgcggc aagcgcccga cagccttcca
      661 gttcctgcaa gaaatcaagc cggaagccgt agcgtacatc accattaaga ccactctggc
      721 ttgcctaacc agtgctgaca atacaaccgt tcaggctgta gcaagcgcaa tcggtcgggc
      781 cattgaggac gaggctcgct tcggtcgtat ccgtgacctt gaagctaagc acttcaagaa
      841 aaacgttgag gaacaactca acaagcgcgt agggcacgtc tacaagaaag catttatgca
      901 agttgtcgag gctgacatgc tctctaaggg tctactcggt ggcgaggcgt ggtcttcgtg
      961 gcataaggaa gactctattc atgtaggagt acgctgcatc gagatgctca ttgagtcaac
     1021 cggaatggtt agcttacacc gccaaaatgc tggcgtagta ggtcaagact ctgagactat
     1081 cgaactcgca cctgaatacg ctgaggctat cgcaacccgt gcaggtgcgc tggctggcat
     1141 ctctccgatg ttccaacctt gcgtagttcc tcctaagccg tggactggca ttactggtgg
     1201 tggctattgg gctaacggtc gtcgtcctct ggcgctggtg cgtactcaca gtaagaaagc
     1261 actgatgcgc tacgaagacg tttacatgcc tgaggtgtac aaagcgatta acattgcgca
     1321 aaacaccgca tggaaaatca acaagaaagt cctagcggtc gccaacgtaa tcaccaagtg
     1381 gaagcattgt ccggtcgagg acatccctgc gattgagcgt gaagaactcc cgatgaaacc
     1441 ggaagacatc gacatgaatc ctgaggctct caccgcgtgg aaacgtgctg ccgctgctgt
     1501 gtaccgcaag gacaaggctc gcaagtctcg ccgtatcagc cttgagttca tgcttgagca
     1561 agccaataag tttgctaacc ataaggccat ctggttccct tacaacatgg actggcgcgg
     1621 tcgtgtttac gctgtgtcaa tgttcaaccc gcaaggtaac gatatgacca aaggactgct
     1681 tacgctggcg aaaggtaaac caatcggtaa ggaaggttac tactggctga aaatccacgg
     1741 tgcaaactgt gcgggtgtcg ataaggttcc gttccctgag cgcatcaagt tcattgagga
     1801 aaaccacgag aacatcatgg cttgcgctaa gtctccactg gagaacactt ggtgggctga
     1861 gcaagattct ccgttctgct tccttgcgtt ctgctttgag tacgctgggg tacagcacca
     1921 cggcctgagc tataactgct cccttccgct ggcgtttgac gggtcttgct ctggcatcca
     1981 gcacttctcc gcgatgctcc gagatgaggt aggtggtcgc gcggttaact tgcttcctag
     2041 tgaaaccgtt caggacatct acgggattgt tgctaagaaa gtcaacgaga ttctacaagc
     2101 agacgcaatc aatgggaccg ataacgaagt agttaccgtg accgatgaga acactggtga
     2161 aatctctgag aaagtcaagc tgggcactaa ggcactggct ggtcaatggc tggcttacgg
     2221 tgttactcgc agtgtgacta agcgttcagt catgacgctg gcttacgggt ccaaagagtt
     2281 cggcttccgt caacaagtgc tggaagatac cattcagcca gctattgatt ccggcaaggg
     2341 tctgatgttc actcagccga atcaggctgc tggatacatg gctaagctga tttgggaatc
     2401 tgtgagcgtg acggtggtag ctgcggttga agcaatgaac tggcttaagt ctgctgctaa
     2461 gctgctggct gctgaggtca aagataagaa gactggagag attcttcgca agcgttgcgc
     2521 tgtgcattgg gtaactcctg atggtttccc tgtgtggcag gaatacaaga agcctattca
     2581 gacgcgcttg aacctgatgt tcctcggtca gttccgctta cagcctacca ttaacaccaa
     2641 caaagatagc gagattgatg cacacaaaca ggagtctggt atcgctccta actttgtaca
     2701 cagccaagac ggtagccacc ttcgtaagac tgtagtgtgg gcacacgaga agtacggaat
     2761 cgaatctttt gcactgattc acgactcctt cggtaccatt ccggctgacg ctgcgaacct
     2821 gttcaaagca gtgcgcgaaa ctatggttga cacatatgag tcttgtgatg tactggctga
     2881 tttctacgac cagttcgctg accagttgca cgagtctcaa ttggacaaaa tgccagcact
     2941 tccggctaaa ggtaacttga acctccgtga catcttagag tcggacttcg cgttcgcggg
     3001 atccccgggt accgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3061 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3121 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3181 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
     3241 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3301 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3361 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3421 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3481 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3541 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3601 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3661 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3721 gtacaagtaa aagcttaatt agctgaccta ctagtcggcc gtctcgagta gaggcatcaa
     3781 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3841 aacgctctcc tgagtaggac aaatccgccg ccctagacct aggggatata ttccgcttcc
     3901 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     3961 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     4021 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     4081 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     4141 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     4201 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     4261 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     4321 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     4381 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     4441 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     4501 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     4561 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt actagatttc
     4621 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt
     4681 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     4741 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc
     4801 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4861 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4921 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4981 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     5041 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     5101 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5161 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5221 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5281 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5341 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5401 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     5461 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5521 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5581 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5641 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     5701 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccga cgtctgtgtg
     5761 gaattgtgag cggataacaa tttcacacag ggccctcgga caccgaggag aatgtcaaga
     5821 ggcgaacaca caacgtcttg gagcgccaga ggaggaacga gctaaaacgg agcttttttg
     5881 ccctgcgtga ccagatcccg gagttggaaa acaat
//
