LOCUS       sequence-147696-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 3. Integration Site 8..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  AUTHORS   Perez-Pinera P, Han N, Cleto S, Cao J, Purcell O, Shah KA, Lee K, 
            Ram R, Lu TK
  TITLE     Synthetic biology and microbioreactor platforms for programmable 
            production of biologics at the point-of-care.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12211. doi: 10.1038/ncomms12211.
  PUBMED    27470089
REFERENCE   2  (bases 1 to 3214)
  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: "2016-07-29"; volume: "7:12211. doi"; pages: " 
            10.1038/ncomms12211"
COMMENT     Sequence Label: PP67
FEATURES             Location/Qualifiers
     source          1..3214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      98..345
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(114..132)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(420..1008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(500..519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1099..1575
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     protein_bind    1637..1684
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        2000..2413
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2420..2467
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2492..3214,1..93)
                     /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             join(2492..3214,1..93)
                     /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(2565..2584)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
       61 cagtttcatt tgatgctcga tgagtttttc taacacatca tgtaattagt tatgtcacgc
      121 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
      181 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
      241 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
      301 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac
      361 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      421 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      481 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      541 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      601 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      661 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      721 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      781 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      841 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      901 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      961 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1021 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1081 atgcatgaga tcagatcttt tttgtagaaa tgtcttggtg tcctcgtcca atcaggtagc
     1141 catctctgaa atatctggct ccgttgcaac tccgaacgac ctgctggcaa cgtaaaattc
     1201 tccggggtaa aacttaaatg tggagtaatg gaaccagaaa cgtctcttcc cttctctctc
     1261 cttccaccgc ccgttaccgt ccctaggaaa ttttactctg ctggagagct tcttctacgg
     1321 cccccttgca gcaatgctct tcccagcatt acgttgcggg taaaacggag gtcgtgtacc
     1381 cgacctagca gcccagggat ggaaaagtcc cggccgtcgc tggcaataat agcgggcgga
     1441 cgcatgtcat gagattattg gaaaccacca gaatcgaata taaaaggcga acacctttcc
     1501 caattttggt ttctcctgac ccaaagactt taaatttaat ttatttgtcc ctatttcaat
     1561 caattgaaca actatttcgg ctggacggcg acgtaaacgg ccacaagttt atggccgtga
     1621 tgacctgtgt cttcgtggtt tgtctggtca accaccgcgg tctcagtggt gtacggtaca
     1681 aacccaaagc agcacgacac ggcaactaca agacccgcgc cgagggcatg ttccccaaag
     1741 cgataccact tgaagcagtg gtactgcttg tgggtacact ctgcgggtgt gaagttcgag
     1801 ggcgacaccc tggtgaaccg catcgagctg aagggcatct tccaactcgc ttaattgcga
     1861 gtttttattt cgtttatttc aattaaggta actaaaaaac tccttttaca catgaagcag
     1921 cacgacttct tcaagtccgc catgcccgaa aaacgcctct tcagagtaca gaagattaag
     1981 tgagaccttc gtttgtgcgg atcccccaca caccatagct tcaaaatgtt tctactcctt
     2041 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     2101 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     2161 aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     2221 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc
     2281 gatgacctcc cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt
     2341 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc
     2401 aatctaatct aagggcggtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2461 atacgacaag gtgaggaact aaaccatggt aatgagccat attcaacggg aaacgtcttg
     2521 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2581 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     2641 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2701 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2761 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     2821 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2881 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2941 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3001 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3061 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3121 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3181 catcctatgg aactgcctcg gtgagttttc tcct
//
