LOCUS       sequence-12664-e        6163 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 6163)
  AUTHORS   Dunlop MJ, Cox RS, Levine JH, Murray RM, Elowitz MB
  TITLE     Regulatory activity revealed by dynamic correlations in gene 
            expression noise.
  JOURNAL   Nat Genet. 2008 Dec . 40(12):1493-8.
  PUBMED    19029898
REFERENCE   2  (bases 1 to 6163)
  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 Genet. 
            2008 Dec . 40(12):1493-8."
COMMENT     Sequence Label: pNNS2-ySEc
FEATURES             Location/Qualifiers
     source          1..6163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             444..455
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(1196..1229)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      1238..1309
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1325..1352
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2131..3081)
                     /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      3129..3351
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3843..3937)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3968..4762)
                     /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(4079..4098)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4689..4708
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4941..4959
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4993..5079)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5109..5822)
                     /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
                     FICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             5831..5842
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ggatccgatg cgcctgcctg tattacccat cgtcttgagt tcgtgccgga cggtggactc
       61 tatcgctatg tgctgacaaa aatatattga gtcggtagtg ctgaccttgc cggaggcggc
      121 cttagcaccc tctccggcca acggttcgac gcatgcaggc atgaaaccgc gtcttttttc
      181 agataaaaag cgcaatcatt cataaaccct ctgttttata atcacttaat cgcgcataaa
      241 aaacggctaa attcttgtgt aaacgattcc actaatttat tccatgtcac acttttcgca
      301 tctttgttat gctatggtta tttcatacca taagcctaat ggagcgaatt atgagagttc
      361 tggttaccgg tggtagcggt tacattggaa gtcatacctg tgtgcaatta ctgcaaaacg
      421 gtcatgatgc tcgagttaac tttaaagagg agaaaggtac cgcatgacta gcaaaagaag
      481 caaaggtgaa gaactgttca ctggtgttgt tccaattctg gttgaactgg atggtgatgt
      541 taatggtcac aaattttctg tctctggtga gggtgaaggt gatgcaacct acggtaaact
      601 gaccctgaaa tttatttgca ctactggtaa actgcctgtt ccgtggccaa ccctggtcac
      661 tactctgact tggggtgttc aatgctttgc tcgttaccca gatcacatga aacagcatga
      721 ctttttcaag tctgccatgc cggaaggtta tgttcaggaa cgtactatct ttttcaaaga
      781 tgacggtaac tacaagaccc gtgctgaagt caagtttgaa ggtgataccc tggttaatcg
      841 tatcgagctg aaaggtattg attttaaaga agatggtaac attctgggtc acaaactgga
      901 atacaacgct atttctgata atgtatacat cactgctgac aaacaaaaga atggtatcaa
      961 agctaatttc aaaattcgtc acaacattga agatggtagc gttcaactgg cagaccatta
     1021 tcaacaaaat actccaattg gcgatggccc tgtcctgctg ccagacaacc attacctgtc
     1081 cacccaatct cgtctgtcta aagatccgaa cgaaaagcgc gatcacatgg tcctgctgga
     1141 gtttgtaacc gctgctggta ttaccctggg catggatgaa ctgtataaat aatagataac
     1201 ttcgtatagc atacattata cgaagttatc caggcatcaa ataaaacgaa aggctcagtc
     1261 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     1321 cactggctca ccttcgggtg ggcctttctg cgtttaacag ctattgtaga taagtgggga
     1381 tactacgggt ataaaaaaac ccgccggggc gggttttttt acgttgcttc agagctgatg
     1441 tggtggagat cgaatttcaa aggttagttt tttcatgact tccctctccc ccaaataaaa
     1501 aggcctgctg gcgattacca gccagcaggc ctagtattaa ccgtgtcctt ctcgatttga
     1561 ataaataaaa acagccgttg ccagaaagag gcacggctgt ttttattttt ctagggtacg
     1621 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     1681 tccggcttca ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc
     1741 cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc
     1801 gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca
     1861 atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc
     1921 tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttaaatgc accaaaaact
     1981 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     2041 tttccctttg atatctaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     2101 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     2161 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatgctt
     2221 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     2281 agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag gaatctgatg taatggttgt
     2341 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     2401 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     2461 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     2521 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     2581 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     2641 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     2701 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     2761 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     2821 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     2881 ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac
     2941 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     3001 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     3061 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     3121 attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg
     3181 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     3241 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     3301 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     3361 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     3421 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     3481 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     3541 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     3601 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     3661 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     3721 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     3781 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt
     3841 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     3901 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga
     3961 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     4021 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4081 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4141 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4201 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     4261 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4321 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4381 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4441 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4501 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     4561 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4621 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4681 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4741 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg
     4801 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     4861 tcccaacctt accagagggc gccccagctg gcaattccga cgtctaagaa accattatta
     4921 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacggatctt
     4981 ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     5041 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctaga ctcagctaat
     5101 taagcttatt tgtatagttc atccatgcca tgtgtaatcc cagcagctgt tacaaactca
     5161 agaaggacca tgtggtctct cttttcgttg ggatctttcg aaagggcaga ttgataggac
     5221 aggtaatggt tgtctggtaa aaggacaggg ccatcgccaa ttggagtatt ttgttgataa
     5281 tggtctgcta gttgaacgct tccatcttca atgttgtgtc taattttgaa gttaactttg
     5341 attccattct tttgtttgtc tgccatgatg tatacattgt gtgagttata gttgtattcc
     5401 aatttgtgtc caagaatgtt tccatcttct ttaaaatcaa taccttttaa ctcgattcta
     5461 ttaacaaggg tatcaccttc aaacttgact tcagcacgtg tcttgtagtt cccgtcatct
     5521 ttgaaaaata tagttctttc ctgtacataa ccttcgggca tggcactctt gaaaaagtca
     5581 tgccgtttca tatgatctgg gtatctagca aagcatttta gaccataacc gaaagtagtg
     5641 acaagtgttg gccatggaac aggtagtttt ccagtagtgc aaataaattt aagggtaagt
     5701 tttccgtatg ttgcatcacc ttcaccctct ccactgacag aaaatttgtg cccattaaca
     5761 tcaccatcta attcaacaag aattgggaca actccagtga aaagttcttc tcctttacgc
     5821 atgcggtacc tttctcctct ttaatgaatt gtcgactatt gagcacccgg gaaaccgttg
     5881 ccacagagac gccagcctga cgcgctacat cacgaatggt gatcatattc actacctgtt
     5941 atgagattgc agtaaatgac tgcttgctgg cggctatttt gtcagcactg aatacaggac
     6001 ttcgtgaatc gagtcacagc aatggaaacg gttacagccg ttttgttaac gaatgtgatc
     6061 cagatcgtta tcttgatgtt ttagatattg ccggtccggc agcacgcaaa gttaactgac
     6121 gccagagcca ttccaccggc cctggcggaa gtaacgcagc cag
//
