LOCUS       sequence-466455-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 0 5 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3210)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 3210)
  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"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl0-G-iRFP720
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     regulatory      841..850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             847..1797
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     primer_bind     complement(1837..1855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1923..2027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2249..3040
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaggt
      841 gccaccatgg cggaaggatc cgtcgccagg cagcctgacc tcttgacctg cgacgatgag
      901 ccgatccata tccccggtgc catccaaccg catggactgc tgctcgccct cgccgccgac
      961 atgacgatcg ttgccggcag cgacaacctt cccgaactca ccggactggc gatcggcgcc
     1021 ctgatcggcc gctctgcggc cgatgtcttc gactcggaga cgcacaaccg tctgacgatc
     1081 gccttggccg agcccggggc ggccgtcgga gcaccgatca ctgtcggctt cacgatgcga
     1141 aaggacgcag gcttcatcgg ctcctggcat cgccatgatc agctcatctt cctcgagctc
     1201 gagcctcccc agcgggacgt cgccgagccg caggcgttct tccgccgcac caacagcgcc
     1261 atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct gcgccgccgc ggcgcaagag
     1321 gtgcggaaga ttaccggctt cgatcgggtg atgatctatc gcttcgcctc cgacttcagc
     1381 gggtccgtga tcgcagagga tcggtgcgcc gaggtcgagt caaaactagg cctgcactat
     1441 cctgcctcat tcatcccggc gcaggcccgt cggctctata ccatcaaccc ggtacggatc
     1501 attcccgata tcaattatcg gccggtgccg gtcaccccag acctcaatcc ggtcaccggg
     1561 cggccgattg atcttagctt cgccatcctg cgcagcgtct cgcccaacca tctggagttc
     1621 atgcgcaaca taggcatgca cggcacgatg tcgatctcga ttttgcgcgg cgagcgactg
     1681 tggggattga tcgtttgcca tcaccgaacg ccgtactacg tcgatctcga tggccgccaa
     1741 gcctgcgagc tagtcgccca ggttctggcc tggcagatcg gcgtgatgga agagtaagct
     1801 ttgagaccac gaagtggctc ttcagtggac gaaagggcct cgtgatacgc ctatttttat
     1861 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1921 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1981 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg cgctcacgca
     2041 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     2101 cttgttatga ctgttttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     2161 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     2221 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     2281 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     2341 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagcgata ttgatttgct
     2401 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     2461 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     2521 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     2581 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     2641 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     2701 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     2761 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     2821 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     2881 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     2941 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     3001 ttacgtaaaa ggcgagatca ccaaggtagt cggcaaataa ctgtcagacc aagtttactc
     3061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3181 agaccccgta gaaaagatca aaggatcttc
//
