LOCUS       sequence-147694-        3468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 3. Integration Site 6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3468)
  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 3468)
  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: PP79
FEATURES             Location/Qualifiers
     source          1..3468
                     /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    2227..2274
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        2590..3003
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3010..3057
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3082..3468,1..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(3082..3468,1..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(3155..3174)
                     /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 atctaggcgt
     1441 agtcttgtcg aatctacccc aggtactggt gctagtagag ccagaacttc ccacagtatt
     1501 gtagtagtaa gagtaaggtg tagtcttgtc aaatcttccc cagggtgtaa agatggacac
     1561 cgacatgtcg gctatggtaa ttgtgactac atgggcatcg gtggctgttg aatcggcggt
     1621 cggagtgacg gtgttgattg caaacaagct caccaagaaa ctggcaaatt gcatcttgtg
     1681 ttgtttgatg ttttaagtca gaattcggtg gaaacttaag ggtccgcata ataaataata
     1741 cagaaaaaaa aagaaagcaa ccagaaaggg aaataaagtt caaaaaattc taagtgttaa
     1801 caggaatata ttaatgagtc tatggtatct ctctacaaga ccatcagtaa accgaggcca
     1861 gcagcaccaa ctaaaccgaa ctgggcacca accgaggcag ccaaaccttc ataactggaa
     1921 gtggtagtgt tggagtggtt agagctgttc gaaccgttaa taacaacttc tctgtgaaca
     1981 acaccgtcgt acgagaatat ggtggtagtg gtaggatctg gagttttgtt aaatcttcat
     2041 gagattattg gaaaccacca gaatcgaata taaaaggcga acacctttcc caattttggt
     2101 ttctcctgac ccaaagactt taaatttaat ttatttgtcc ctatttcaat caattgaaca
     2161 actatttcgg ctggacggcg acgtaaacgg ccacaagttt atggccgtga tgacctgtgt
     2221 cttcgtggtt tgtctggtca accaccgcgg tctcagtggt gtacggtaca aacccaaagc
     2281 agcacgacac ggcaactaca agacccgcgc cgagggcatg ttccccaaag cgataccact
     2341 tgaagcagtg gtactgcttg tgggtacact ctgcgggtgt gaagttcgag ggcgacaccc
     2401 tggtgaaccg catcgagctg aagggcatct tccaactcgc ttaattgcga gtttttattt
     2461 cgtttatttc aattaaggta actaaaaaac tccttttaca catgaagcag cacgacttct
     2521 tcaagtccgc catgcccgaa aaacgcctct tcagagtaca gaagattaag tgagaccttc
     2581 gtttgtgcgg atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt
     2641 ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata
     2701 ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga
     2761 aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt
     2821 atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc
     2881 cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt
     2941 gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct
     3001 aagggcggtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3061 gtgaggaact aaaccatggt aatgagccat attcaacggg aaacgtcttg ctctaggccg
     3121 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     3181 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     3241 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     3301 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     3361 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     3421 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgc
//
