LOCUS       sequence-335352-        3405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23101 driving PP7 repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3405)
  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 3405)
  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: pSB207
FEATURES             Location/Qualifiers
     source          1..3405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(369..1082,1083..1085,1086..1088))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(414..435)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     763..782
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1022..1043
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_RNA        complement(1138..1187)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        complement(1194..1228)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     primer_bind     complement(1390..1409)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1401..2216
                     /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             1401..2216
                     /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(1474..1493)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2305..2850
                     /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     2751..2770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3215..3234
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatctaatga acccgggaat actgcagcca gcgagcccat atgctctgct ggtttctaga
       61 gtcgacctgc agggagcgac gccatatcgt ctgctccttt ctaggcaatt aggtacctta
      121 ggatctaatg aacccgggaa tactgcagcc agcgagccca tatgctctgc tggtttctag
      181 agtcgacctg cagggagcga cgccatatcg tctgctcctt tctaggcaat taggtacctt
      241 aggatctaat gaacccggga atactgcagc cagcgagccc atatgctctg ctggtttcta
      301 gagtcgacct gcagggagcg acgccatatc gtctgctcct ttctaggcaa ttaggtacct
      361 taggatcctt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
      421 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag
      481 ctaaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgattggggt gttctgctgg
      541 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc
      601 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac
      661 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
      721 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
      781 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
      841 tcgtgcagct tcatgtggtc ggggtagcgg gcgaagcatt gcaggccgta gccgaaggtg
      901 gtcacgaggg tgggccaggg cacgggcagc ttgcctgtgg tgcagatgaa cttcagggtc
      961 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     1021 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     1081 ctcaccatct agtatttccc ctctttctct agtattaaac aaaattattt gtagaggctg
     1141 tttcgtcctc acggactcat cagaccggaa agcacatccg gtgacagctt caggctagca
     1201 taatacctag gactgagcta gctgtaaaag ttagggactt atcaagcggg agaccagaaa
     1261 caaaaaaagg ccccccgtta gggaggcctt caataattgg ttgtgtctca aaatctctga
     1321 tgtacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1381 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tccaggccgc
     1441 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1501 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     1561 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1621 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1681 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     1741 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1801 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1861 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1921 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     1981 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2041 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2101 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2161 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaaggta
     2221 actgtcagac caagtttact catatatact ttagattgat ttttcgttcc actgagcgtc
     2281 agacccctta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     2341 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     2401 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     2461 gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg
     2521 ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg
     2581 cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct
     2641 acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca
     2701 ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     2821 tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc
     2881 tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg
     2941 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc
     3001 tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac
     3061 ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgctga
     3121 ggtctgcctc gtgcagcgag tcagtgagcg aggaagcacc tcagataaaa tatttgctca
     3181 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     3241 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcggac
     3301 caaaacgaaa aaaggccccc ctttcgggag gcctcttttc tggaatttgg taccgagctg
     3361 cagcgggcgc tactagtata gcggccgcga attcgccctt cgcga
//
