LOCUS       sequence-174950-        2660 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 2660)
  AUTHORS   Basu S, Mehreja R, Thiberge S, Chen MT, Weiss R
  TITLE     Spatiotemporal control of gene expression with pulse-generating 
            networks.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Apr 27. 101(17):6355-60.
  PUBMED    15096621
REFERENCE   2  (bases 1 to 2660)
  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: "Proc Natl 
            Acad Sci U S A. 2004 Apr 27. 101(17):6355-60."
COMMENT     Sequence Label: pPSSUB-101
FEATURES             Location/Qualifiers
     source          1..2660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(175..269)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(290..949)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     864..883
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(950..1052)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1118..1136
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             1268..1279
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1286..1999
                     /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"
     CDS             1286..1999
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1316..1344)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1939..1959
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      complement(join(2159..2660,1..87))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2239..2258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
       61 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      121 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactagtgc ttggattctc
      181 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
      241 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
      301 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
      361 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
      421 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
      481 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
      541 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
      601 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
      661 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
      721 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
      781 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
      841 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
      901 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
      961 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     1021 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccag atatcgacgt
     1081 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1141 tcgtcttcat ctcttgtcac actattgtat cgctgggaat acaattactt aacataagca
     1201 cctgtaggat cgtacaggtt tacgcaagaa aatggtttgt tatagtcgaa tacatctggc
     1261 ggtgataaaa gaggagaaat taagcatgcg taaaggagaa gaacttttca ctggagttgt
     1321 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1381 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     1441 actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc aatgctttgc
     1501 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
     1561 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     1621 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     1681 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
     1741 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
     1801 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     1861 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     1921 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     1981 catggatgaa ctatacaaaa ggcctgcagc aaacgacgaa aactacgctt tagtagctta
     2041 atacagtaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2101 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2581 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2641 tcggaaaaag agttggtagc
//
