LOCUS       sequence-147457-        4385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output plasmid: PPhlF-PAmtR-YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4385)
  AUTHORS   Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski
            EA, Ross D, Densmore D, Voigt CA
  TITLE     Genetic circuit design automation.
  JOURNAL   Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341.
  PUBMED    27034378
REFERENCE   2  (bases 1 to 4385)
  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: "Science."; 
            date: "2016-04-1"; volume: "352(6281):aac7341. doi"; pages: " 
            10.1126/science.aac7341"
COMMENT     Sequence Label: pAN4036
FEATURES             Location/Qualifiers
     source          1..4385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..792
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        1088..1137
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(1187..1189,1190..1192,1193..1906)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1232..1253)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1493..1512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1840..1861
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2109..2203
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2695..2917
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2965..3915
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
       61 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
      121 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
      181 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
      241 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
      301 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
      361 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
      421 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
      481 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
      541 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
      601 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
      661 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
      721 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
      781 gtcggcaaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
      841 ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga actcggtacc
      901 aaaccaatta ttgaagacgc tgaaaagcgt ctttttttgt ttcggtccgc ttcgacgtac
      961 ggtggaatct gattcgttac caattgacat gatacgaaac gtaccgtatc gttaaggtct
     1021 tgtccaacca aatgattcgt taccaattga cagtttctat cgatctatag ataatgctag
     1081 cctgaagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct
     1141 ctacaaataa ttttgtttaa tactagagaa agaggggaaa tactagatgg tgagcaaggg
     1201 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1261 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1321 gaagttcatc tgcaccacag gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     1381 cggctacggc ctgcaatgct tcgcccgcta ccccgaccac atgaagctgc acgacttctt
     1441 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1501 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1561 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1621 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1681 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1741 gaacacccca atcggcgacg gccccgtgct gctgcccgac aaccactacc ttagctacca
     1801 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1861 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaactcg gtaccaaatt
     1921 ccagaaaaga ggcctcccga aaggggggcc tttttcgttt tggtccggct gaagaccagc
     1981 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaac tgtcagacca agtttacgag
     2101 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2161 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagggacag
     2221 taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa
     2281 tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct
     2341 gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc
     2401 ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat aaaaggcgcc
     2461 tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac
     2521 gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt cagcgatttg
     2581 cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt ttgtagagga
     2641 gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag tagtgagtta
     2701 tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat tctataaatt
     2761 ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt
     2821 ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat acccacaact
     2881 caaaggaaaa ggacatgtaa ttatcattga ctagcccatc tcaattggta tagtgattaa
     2941 aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc aaatgaacta
     3001 gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg ctgtgtggca
     3061 ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc gttcacttat
     3121 aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt attagctaaa
     3181 gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt taaaggcttt
     3241 gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga attagttttt
     3301 agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata taatctggaa
     3361 catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt attaaaagaa
     3421 ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga atttaagttc
     3481 atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa
     3541 ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga taagcgaggc
     3601 cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga tctcgtaacc
     3661 gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac cattacatca
     3721 gattcctacc tacgtaacgg actaagaaaa acactacacg atgctttaac tgcaaaaatt
     3781 cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagca tgatctcaat
     3841 ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat actggctaaa
     3901 tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat caagactaac
     3961 aaacaaaagt agaacaactg ttcaccgtta gatatcaaag ggaaaactgt ccatatgcac
     4021 agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt ttggtgcatt
     4081 taaagctgtt caccatgaac agatcgacaa tgtaacgatt atcaaaaagg atcttcacct
     4141 agatcctttt aaattaaaaa tgaagcttcg ttgtgtctca aaatctctga tgttacattg
     4201 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4261 caaggggtgt ttactagagg ttgatcgggc acgtaagagg ttccaacttt caccataatg
     4321 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     4381 ctaaa
//
