LOCUS       sequence-147697-        3763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 3. Integration Site 9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3763)
  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 3763)
  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: PP151
FEATURES             Location/Qualifiers
     source          1..3763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      414..661
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(430..448)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(736..1324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(816..835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1415..1448)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    2502..2549
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        2865..3278
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3285..3332
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3357..3763,1..409)
                     /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(3357..3763,1..409)
                     /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(3430..3449)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
       61 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
      121 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
      181 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
      241 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
      301 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
      361 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaac acatcatgta
      421 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
      481 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
      541 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
      601 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
      661 caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1381 ttaagggatt ttggtcatgc atgagatcag atctgaagtt cctatacttt ctagagaata
     1441 ggaacttcaa gcttaaaacc atgttcgcaa gctgaaagaa catattctga aaggagacat
     1501 cttccaagct gttccctctc aaagggtagc caggccgacc tcattgcacc ctttcaacat
     1561 ctatcgtcat ttgagaactg tcaatccttc tccatacatg ttctatattg actatctaga
     1621 cttccaagtt gttggtgctt cacctgaatt actagttaaa tccgacaaca acaacaaaat
     1681 catcacacat cctattgctg gaactcttcc cagaggtaaa actatcgaag aggacgacaa
     1741 ttatgctaag caattgaagt cgtctttgaa agacagggcc gagcacgtca tgctggtaga
     1801 tttggccaga aatgatatta accgtgtgtg tgagcccacc agtaccacgg ttgatcgttt
     1861 attgactgtg gagagatttt ctcatgtgat gcatcttgtg tcagaagtca gtggaacatt
     1921 gagaccaaac aagactcgct tcgatgcttt cagatccact ttcccagcag gtaccgtctc
     1981 cggtgctccg aaggtaagag caatgcaact cataggagaa ttggaaggag aaaagagagg
     2041 tgtttatgcg ggggccgtag gacactggtc gtacgatgga aaatcgatgg acacatgtat
     2101 tgccttaaga acaatggtcg tcaaggacgg tgtcgcttac cttcaagccg gaggtggaat
     2161 tgtctacgat tctgacccct atgacgagta catcgaaacc atgaacaaaa tgagatccaa
     2221 caataacacc atcttggagg ctgagaaaat ctggaccgat aggttggcca gagacgagaa
     2281 tcaaagtgaa tccgaagaaa acgatcaatg atcatgagat tattggaaac caccagaatc
     2341 gaatataaaa ggcgaacacc tttcccaatt ttggtttctc ctgacccaaa gactttaaat
     2401 ttaatttatt tgtccctatt tcaatcaatt gaacaactat ttcggctgga cggcgacgta
     2461 aacggccaca agtttatggc cgtgatgacc tgtgtcttcg tggtttgtct ggtcaaccac
     2521 cgcggtctca gtggtgtacg gtacaaaccc aaagcagcac gacacggcaa ctacaagacc
     2581 cgcgccgagg gcatgttccc caaagcgata ccacttgaag cagtggtact gcttgtgggt
     2641 acactctgcg ggtgtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2701 catcttccaa ctcgcttaat tgcgagtttt tatttcgttt atttcaatta aggtaactaa
     2761 aaaactcctt ttacacatga agcagcacga cttcttcaag tccgccatgc ccgaaaaacg
     2821 cctcttcaga gtacagaaga ttaagtgaga ccttcgtttg tgcggatccc ccacacacca
     2881 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     2941 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     3001 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     3061 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     3121 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     3181 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     3241 ttgctcatta gaaagaaagc atagcaatct aatctaaggg cggtgttgac aattaatcat
     3301 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggtaatga
     3361 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     3421 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3481 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3541 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3601 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3661 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3721 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgt
//
