LOCUS       sequence-335356-        3667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p15A backbone with PhlF binding sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3667)
  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 3667)
  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: pSB219
FEATURES             Location/Qualifiers
     source          1..3667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..44)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        89..123
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        131..180
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(223..225,226..939)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             223..936
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(253..281)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     876..896
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      956..1027
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1043..1070
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1518..1538
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1539..1596
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      1709..2254
                     /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     2155..2174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2613..3404,3405..3473))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3236..3255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3474..3578)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3646..3664
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
       61 ttctagaggg aggtgtcttc ggcgcgcctt tacggctagc tcagtcctag gtactatgct
      121 agcaaggtag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag
      181 cctctacaaa taattttgtt taatcacaca ggaaagtact agatgagcaa aggagaagaa
      241 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      301 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt
      361 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat
      421 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt
      481 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac
      541 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag
      601 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac
      661 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
      721 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact
      781 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc
      841 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
      901 gctgggatta cacatggcat ggatgagctc tacaaataat actagagcca ggcatcaaat
      961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1021 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatagaag
     1081 acatcgctta ctagagtaca tgatacgaaa cgtaccgtat cgttaaggtg catgatacga
     1141 aacgtaccgt atcgttaagg ttgtatgata cgaaacgtac cgtatcgtta aggtgcatga
     1201 tacgaaacgt accgtatcgt taaggtctga tgatacgaaa cgtaccgtat cgttaaggta
     1261 aatgatacga aacgtaccgt atcgttaagg tgcatgatac gaaacgtacc gtatcgttaa
     1321 ggtaggtatg atctaactac tagagtacat gatacgaaac gtaccgtatc gttaaggtca
     1381 tgatacgaaa cgtaccgtat cgttaaggtt gtatgatacg aaacgtaccg tatcgttaag
     1441 gtgcatgata cgaaacgtac cgtatcgtta aggtctgatg atacgaaacg taccgtatcg
     1501 ttaaggtatg atctaactac tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg
     1561 tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc
     1621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1681 aaggcggtaa tttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     1741 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     1801 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     1861 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     1921 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     1981 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     2041 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     2101 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     2221 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     2281 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     2341 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     2401 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     2461 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctacga
     2521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     2761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tataacccac tcgtgcaccc
     3361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3601 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3661 aggcaga
//
