LOCUS       sequence-147690-        3469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 3. Integration Site 3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3469)
  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 3469)
  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: PP76
FEATURES             Location/Qualifiers
     source          1..3469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      434..681
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(450..468)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(756..1344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(836..855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2228..2275
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        2591..3004
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3011..3058
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3083..3469,1..429)
                     /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(3083..3469,1..429)
                     /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(3156..3175)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
       61 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
      121 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
      181 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
      241 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
      301 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
      361 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
      421 tttttctaac acatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
      481 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
      541 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
      601 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
      661 cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag
      721 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      781 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      841 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      901 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      961 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1021 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1081 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1141 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1201 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1261 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1321 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1381 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc atgagatcag atctcagtag
     1441 ttggttttgt ttcaaactgc ttatagctta ctaaaggaat cgctgtattc agtcaagtta
     1501 tactgtcgta ttccagagaa gtagatttct gtgaacttta tggagctgat gagttgaatt
     1561 tgccggggcg tgctcttttt tcatttatct tgtagttcat tttctacgtc gggttgttac
     1621 agttatcgag atggagtagc aacagtgaga tagaacttct gccctattga attgctttct
     1681 tctataagtc ttctttcgta aactgttcct gcagtttttg cgtatagtaa atgaagactt
     1741 ttactggtca cacgaattct cagattttga ttgggtgccc aatggtagat ggttgctgga
     1801 cgggtttgtt aactatattt catgtcgaac caatatggac atatgtcgga ggcacccatg
     1861 atgccactaa aagtagggcc attttttttc gtaagtgtca cccaaagaaa agccattggc
     1921 tattactaac agagtaattt ttgcatttta actacatggt gtcctttact ccaacccggg
     1981 atactaatcg tcaagaactg gatagaaaga gaaatgtagc actagccaac tttaaaatca
     2041 tgagattatt ggaaaccacc agaatcgaat ataaaaggcg aacacctttc ccaattttgg
     2101 tttctcctga cccaaagact ttaaatttaa tttatttgtc cctatttcaa tcaattgaac
     2161 aactatttcg gctggacggc gacgtaaacg gccacaagtt tatggccgtg atgacctgtg
     2221 tcttcgtggt ttgtctggtc aaccaccgcg gtctcagtgg tgtacggtac aaacccaaag
     2281 cagcacgaca cggcaactac aagacccgcg ccgagggcat gttccccaaa gcgataccac
     2341 ttgaagcagt ggtactgctt gtgggtacac tctgcgggtg tgaagttcga gggcgacacc
     2401 ctggtgaacc gcatcgagct gaagggcatc ttccaactcg cttaattgcg agtttttatt
     2461 tcgtttattt caattaaggt aactaaaaaa ctccttttac acatgaagca gcacgacttc
     2521 ttcaagtccg ccatgcccga aaaacgcctc ttcagagtac agaagattaa gtgagacctt
     2581 cgtttgtgcg gatcccccac acaccatagc ttcaaaatgt ttctactcct tttttactct
     2641 tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat
     2701 actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg
     2761 aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt
     2821 tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt cgatgacctc
     2881 ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct
     2941 tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag caatctaatc
     3001 taagggcggt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3061 ggtgaggaac taaaccatgg taatgagcca tattcaacgg gaaacgtctt gctctaggcc
     3121 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     3181 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     3241 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3301 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3361 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     3421 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgc
//
