LOCUS       sequence-335351-        3422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptac driving PP7 repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3422)
  AUTHORS   Shao B, Rammohan J, Anderson DA, Alperovich N, Ross D, Voigt CA
  TITLE     Single-cell measurement of plasmid copy number and promoter 
            activity.
  JOURNAL   Nat Commun. 2021 Mar 5;12(1):1475. doi: 10.1038/s41467-021-21734-y.
  PUBMED    33674569
REFERENCE   2  (bases 1 to 3422)
  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 
            Commun."; date: "2021-03-5"; volume: "12(1):1475. doi"; pages: " 
            10.1038/s41467-021-21734-y"
COMMENT     Sequence Label: pSB208
FEATURES             Location/Qualifiers
     source          1..3422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(353..1066,1067..1069,1070..1072))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(398..419)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     747..766
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1006..1027
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_RNA        complement(1122..1171)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    1174..1193
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(join(1199..1205,1206..1221,1222..1227))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     primer_bind     complement(1391..1410)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1402..2217
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1402..2217
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1475..1494)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2306..2851
                     /direction=RIGHT
                     /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     2752..2771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3216..3235
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gaatactgca gccagcgagc ccatatgctc tgctggtttc tagagtcgac ctgcagggag
       61 cgacgccata tcgtctgctc ctttctaggc aattaggtac cttaggatct aatgaacccg
      121 ggaatactgc agccagcgag cccatatgct ctgctggttt ctagagtcga cctgcaggga
      181 gcgacgccat atcgtctgct cctttctagg caattaggta ccttaggatc taatgaaccc
      241 gggaatactg cagccagcga gcccatatgc tctgctggtt tctagagtcg acctgcaggg
      301 agcgacgcca tatcgtctgc tcctttctag gcaattaggt accttaggat ccttacttgt
      361 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
      421 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctaagg tagtggttgt
      481 cgggcagcag cacggggccg tcgccgattg gggtgttctg ctggtagtgg tcggcgagct
      541 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
      601 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
      661 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
      721 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
      781 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc agcttcatgt
      841 ggtcggggta gcgggcgaag cattgcaggc cgtagccgaa ggtggtcacg agggtgggcc
      901 agggcacggg cagcttgcct gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
      961 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1021 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atctagtatt
     1081 tcccctcttt ctctagtatt aaacaaaatt atttgtagag gctgtttcgt cctcacggac
     1141 tcatcagacc ggaaagcaca tccggtgaca gctaattgtg agcgctcaca attccacaca
     1201 ttatacgagc cgatgattaa ttgtcaacac agccaacgat cgttaagcgg gagaccagaa
     1261 acaaaaaaag gccccccgtt agggaggcct tcaataattg gttgtgtctc aaaatctctg
     1321 atgtacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1381 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctccaggccg
     1441 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1501 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     1561 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1621 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1681 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     1741 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1801 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1861 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1921 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     1981 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2041 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2101 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2161 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaaggt
     2221 aactgtcaga ccaagtttac tcatatatac tttagattga tttttcgttc cactgagcgt
     2281 cagacccctt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     2341 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     2401 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     2461 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     2521 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     2581 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     2641 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     2701 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     2761 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     2821 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     2881 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     2941 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     3001 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     3061 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg
     3121 aggtctgcct cgtgcagcga gtcagtgagc gaggaagcac ctcagataaa atatttgctc
     3181 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     3241 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgga
     3301 ccaaaacgaa aaaaggcccc cctttcggga ggcctctttt ctggaatttg gtaccgagct
     3361 gcagcgggcg ctactagtat agcggccgcg aattcgccct tcgcgagatc taatgaaccc
     3421 gg
//
