LOCUS       sequence-141205-        4311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output plasmid: PBM3R1-YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4311)
  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 4311)
  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: pAN4023
FEATURES             Location/Qualifiers
     source          1..4311
                     /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        1025..1074
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(1124..1126,1127..1129,1130..1843)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1169..1190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1430..1449)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1777..1798
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2035..2129
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2621..2843
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2891..3841
                     /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 ttaatccgcg
      961 tgataggtct gattcgttac caattgacgg aatgaacgtt cattccgata atgctagcct
     1021 gaagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta
     1081 caaataattt tgtttaatac tagagaaaga ggggaaatac tagatggtga gcaagggcga
     1141 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1201 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1261 gttcatctgc accacaggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
     1321 ctacggcctg caatgcttcg cccgctaccc cgaccacatg aagctgcacg acttcttcaa
     1381 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1441 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1501 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1561 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1621 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1681 caccccaatc ggcgacggcc ccgtgctgct gcccgacaac cactacctta gctaccagtc
     1741 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1801 cgccgccggg atcactctcg gcatggacga gctgtacaag taactcggta ccaaattcca
     1861 gaaaagaggc ctcccgaaag gggggccttt tttcgttttg gtccgctcac tcaaaggcgg
     1921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1981 agcaaaaggc caggaaccgt aaaaactgtc agaccaagtt tacgagctcg cttggactcc
     2041 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     2101 gccgccgggc gttttttatt ggtgagaatc caagcactag ggacagtaag acgggtaagc
     2161 ctgttgatga taccgctgcc ttactgggtg cattagccag tctgaatgac ctgtcacggg
     2221 ataatccgaa gtggtcagac tggaaaatca gagggcagga actgctgaac agcaaaaagt
     2281 cagatagcac cacatagcag acccgccata aaacgccctg agaagcccgt gacgggcttt
     2341 tcttgtatta tgggtagttt ccttgcatga atccataaaa ggcgcctgta gtgccattta
     2401 cccccattca ctgccagagc cgtgagcgca gcgaactgaa tgtcacgaaa aagacagcga
     2461 ctcaggtgcc tgatggtcgg agacaaaagg aatattcagc gatttgcccg agcttgcgag
     2521 ggtgctactt aagcctttag ggttttaagg tctgttttgt agaggagcaa acagcgtttg
     2581 cgacatcctt ttgtaatact gcggaactga ctaaagtagt gagttataca cagggctggg
     2641 atctattctt tttatctttt tttattcttt ctttattcta taaattataa ccacttgaat
     2701 ataaacaaaa aaaacacaca aaggtctagc ggaatttaca gagggtctag cagaatttac
     2761 aagttttcca gcaaaggtct agcagaattt acagataccc acaactcaaa ggaaaaggac
     2821 atgtaattat cattgactag cccatctcaa ttggtatagt gattaaaatc acctagacca
     2881 attgagatgt atgtctgaat tagttgtttt caaagcaaat gaactagcga ttagtcgcta
     2941 tgacttaacg gagcatgaaa ccaagctaat tttatgctgt gtggcactac tcaaccccac
     3001 gattgaaaac cctacaagga aagaacggac ggtatcgttc acttataacc aatacgctca
     3061 gatgatgaac atcagtaggg aaaatgctta tggtgtatta gctaaagcaa ccagagagct
     3121 gatgacgaga actgtggaaa tcaggaatcc tttggttaaa ggctttgaga ttttccagtg
     3181 gacaaactat gccaagttct caagcgaaaa attagaatta gtttttagtg aagagatatt
     3241 gccttatctt ttccagttaa aaaaattcat aaaatataat ctggaacatg ttaagtcttt
     3301 tgaaaacaaa tactctatga ggatttatga gtggttatta aaagaactaa cacaaaagaa
     3361 aactcacaag gcaaatatag agattagcct tgatgaattt aagttcatgt taatgcttga
     3421 aaataactac catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga
     3481 tttaaacact tacagcaata tgaaattggt ggttgataag cgaggccgcc cgactgatac
     3541 gttgattttc caagttgaac tagatagaca aatggatctc gtaaccgaac ttgagaacaa
     3601 ccagataaaa atgaatggtg acaaaatacc aacaaccatt acatcagatt cctacctacg
     3661 taacggacta agaaaaacac tacacgatgc tttaactgca aaaattcagc tcaccagttt
     3721 tgaggcaaaa tttttgagtg acatgcaaag taagcatgat ctcaatggtt cgttctcatg
     3781 gctcacgcaa aaacaacgaa ccacactaga gaacatactg gctaaatacg gaaggatctg
     3841 aggttcttat ggctcttgta tctatcagtg aagcatcaag actaacaaac aaaagtagaa
     3901 caactgttca ccgttagata tcaaagggaa aactgtccat atgcacagat gaaaacggtg
     3961 taaaaaagat agatacatca gagcttttac gagtttttgg tgcatttaaa gctgttcacc
     4021 atgaacagat cgacaatgta acgattatca aaaaggatct tcacctagat ccttttaaat
     4081 taaaaatgaa gcttcgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     4141 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgtttac
     4201 tagaggttga tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     4261 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa a
//
