LOCUS       sequence-147701-        4429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Figure 2. Expresses TP901-1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4429)
  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 4429)
  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: PP45
FEATURES             Location/Qualifiers
     source          1..4429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      568..1156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1057..1076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1277..1684
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(3176..3194)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      3206..3395
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3474..3491)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3480..3704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             join(3877..4429,1..242)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(3877..4429,1..242)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3931..3950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
       61 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
      121 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
      181 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
      241 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
      301 tttcacaccg catacaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      421 tcaataatag cacgtgctaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      481 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg gcttttgctg
     1201 gccttttgct cacatgttct tgactcttcg cgatgtacgg gccagatata cgcgttgaca
     1261 ttgattattg actagtccac acaccatagc ttcaaaatgt ttctactcct tttttactct
     1321 tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat
     1381 actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg
     1441 aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt
     1501 tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt cgatgacctc
     1561 ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct
     1621 tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag caatctaatc
     1681 taaggctagc gtttaaacca ccatgacaaa gaaagtggca atctatacca gagtgagcac
     1741 cacaaaccaa gcagaagaag gtttctctat tgatgaacaa atagatcgtt tgaccaaata
     1801 tgccgaagct atgggttggc aagtgtctga tacatataca gatgccggtt tctctggtgc
     1861 taagctggaa cgccctgcta tgcaaagact gattaatgat atcgaaaaca aagcatttga
     1921 tacagtgctg gtatacaaac tggatagact gagcagatcc gtcagagata cactgtatct
     1981 ggtgaaggat gtattcacaa agaataagat tgattttatt tctctcaatg aatccatcga
     2041 tacaagttcc gctatgggtt cactgttcct cacaatcttg tccgccataa acgaatttga
     2101 acgtgaaaac attaaggaaa gaatgactat gggtaagctg ggtcgcgcaa aatctggtaa
     2161 aagcatgatg tggactaaaa cagcatttgg ttactaccac aatagaaaga caggtatcct
     2221 ggaaattgtg ccactgcaag ctactatcgt tgaacaaatc ttcaccgatt acttgtctgg
     2281 tatttccctg actaagctcc gtgataaact caacgaatct ggtcacatcg gtaaggatat
     2341 cccctggtcc tatagaactc tgagacaaac cctggataat cctgtatact gcggttacat
     2401 caagttcaaa gattcattgt tcgaaggtat gcacaagcca atcatcccct acgaaacata
     2461 cctcaaggta caaaaggaac tggaagaacg acaacaacaa acatacgaac gtaacaacaa
     2521 tccccgccca tttcaagcca aatacatgtt gtccggtatg gctagatgcg gttactgcgg
     2581 tgcacccttg aaaatcgttc tcggtcacaa gcgtaaggat ggttctcgaa ctatgaagta
     2641 ccactgcgca aatagattcc cacgaaaaac gaagggtatc actgtataca acgataacaa
     2701 aaagtgcgat tccggtacat atgatctcag caatcttgaa aatacggtta ttgataacct
     2761 tattggtttc caagaaaata atgattccct gctgaagatc atcaatggta acaatcaacc
     2821 catcttggat accagttcat tcaagaaaca aattagtcaa atcgataaaa aaatccaaaa
     2881 gaactcagat ttgtacttga acgatttcat tactatggat gaacttaagg atagaacaga
     2941 tagcctgcaa gcagaaaaaa agctgctcaa ggccaaaatc agtgaaaaca aattcaacga
     3001 ttccacggat gtctttgaac tcgtgaaaac ccaactgggt agcatcccca tcaacgaact
     3061 cagctatgat aacaaaaaaa agatcgttaa caacttggtt agtaaggtcg atgtcaccgc
     3121 cgataacgtc gatatcatat tcaaattcca actggcctga agatctatta gttatgtcac
     3181 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     3241 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     3301 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     3361 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctggagacca
     3421 acatgtgagc aaaaggccag catctagagg gcccgtttaa acccgctgat cagcctcgac
     3481 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3541 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3601 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3661 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttcta ctgggcggtt
     3721 ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag
     3781 ccctgcaaag taaactggat ggctttctcg ccgccaagga tctgatggcg caggggatca
     3841 agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4381 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcg
//
