LOCUS       sequence-147700-        4381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Figure 2. Expresses R4..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4381)
  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 4381)
  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: PP44
FEATURES             Location/Qualifiers
     source          1..4381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      565..1153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1054..1073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1267..1681
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(3125..3143)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      3155..3344
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3423..3440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3429..3653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             join(3826..4381,1..239)
                     /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(3826..4381,1..239)
                     /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(3880..3899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
       61 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
      121 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
      181 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
      241 ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      301 cacaccgcat acaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      361 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      421 ataatagcac gtgctaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
      721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctgggct tttgctggcc
     1201 ttttgctcac atgttcttga ctcttcgcga tgtacgggcc agatatacgc gttgacattg
     1261 attattgact agtcacacac catagcttca aaatgtttct actccttttt tactcttcca
     1321 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     1381 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     1441 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     1501 acgtttcttt ttcttgaaaa tttttttttt ttgatttttt tctctttcga tgacctccca
     1561 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     1621 tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa
     1681 ggctagcgtt taaaccacca tgaacagagg tggtcctaca gtgagagctg atatttacgt
     1741 acgtatcagc ctggatagaa ccggtgaaga actcggtgta gaaagacaag aagaaagctg
     1801 cagagaactc tgcaagtccc tgggtatgga agttggtcaa gtctgggtag ataacgatct
     1861 gtcagcaaca aaaaagaatg tggtgagacc agatttcgaa gccatgattg ccagcaatcc
     1921 tcaagcaatt gtctgttggc acacagatcg tctcatacga gtgactcgtg atctggaacg
     1981 cgtgatcgat ctgggtgtga acgtccatgc agttatggcc ggtcatctgg atctctcaac
     2041 tcccgccggt cgtgctgtgg ctagaactgt tacggcatgg gcaacctatg aaggtgaaca
     2101 aaaggctgaa cgtcaaaaac tcgccaacat ccaaaacgcc cgcgctggta aaccctacac
     2161 tccaggtatt cgccctttcg gttatggtga tgatcatatg acaatcgtaa ccgctgaagc
     2221 tgatgccatt agagatggtg ccaagatgat cctcgatggt tggagtctgt ctgctgttgc
     2281 ccgctattgg gaagaactca agctgcaaag ccccagaagc atggccgccg gtggtaaggg
     2341 ttggtcactc cgcggtgtca aaaaagtgct gacctcacca cgttacgtgg gtcgaagcag
     2401 ctatttgggt gaagtggtcg gtgatgcaca atggccaccc atcctcgatc ccgatgtgta
     2461 ttacggtgtt gtcgccatcc tcaacaaccc cgatcgattc tcaggtggtc cacgaactgg
     2521 tagaacaccc ggtactctgt tggctggtat cgccctgtgt ggtgaatgcg gtaagactgt
     2581 gtcaggtcga ggttatcgtg gtgtcctggt gtatggttgc aaagatactc acacccgcac
     2641 tccccgatca attgcagatg gtcgcgcaag cagcagtact ttggcacgtc ttatgttccc
     2701 cgattttctc cctggtctgc tcgctagtgg tcaagccgaa gatggtcaat cagcagcctc
     2761 caagcactca gaagctcaaa cacttagaga aagacttgat ggtctggcca cagcttacgc
     2821 agaaggtgct atctctctga gtcaaatgac cgcaggttca gaagccctcc gcaagaagct
     2881 ggaagttatc gaagctgatc tcgtcggttc agccggtatt ccacctttcg atccagtcgc
     2941 tggtgtggcc ggtctcatca gcggttggcc caccactcct ctgccaactc gcagagcttg
     3001 ggttgatttc tgccttgttg tgacactgaa cacacaaaag ggtagacacg cctcctccat
     3061 gacagtggat gatcacgtta ctattgaatg gcgagatgtc gccgaatgaa gatctattag
     3121 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     3181 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     3241 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     3301 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
     3361 tggagaccaa catgtgagca aaaggccagc atctagaggg cccgtttaaa cccgctgatc
     3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctac
     3661 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     3721 gttgggaagc cctgcaaagt aaactggatg gctttctcgc cgccaaggat ctgatggcgc
     3781 aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4381 g
//
