LOCUS       sequence-348366-        4599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of an mRNA encoding Firefly luciferase with A[62]
            tail..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4599)
  AUTHORS   Rojas-Duran MF, Gilbert WV
  TITLE     Alternative transcription start site selection leads to large 
            differences in translation activity in yeast.
  JOURNAL   RNA. 2012 Dec;18(12):2299-305. doi: 10.1261/rna.035865.112. Epub 
            2012 Oct 25.
  PUBMED    23105001
REFERENCE   2  (bases 1 to 4599)
  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: 
            "10.1261/rna.035865.112"; journalName: "RNA"; date: "2012-12"; 
            volume: "18"; issue: "12"; pages: "2299-305"
COMMENT     Sequence Label: wg186Fluc
FEATURES             Location/Qualifiers
     source          1..4599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             171..1823
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(237..255)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(1826..1842)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1826..1842)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1949..1966)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1949..1965)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1958..1980)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2106..2561)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2243..2264)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2455..2474
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2588..2692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2693..2761,2762..3553)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2911..2930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3724..4312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4213..4232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4466..4483
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcattaata cgactcacta taggcctaag cttgtcgacc atggaagacg
      181 ccaaaaacat aaagaaaggc ccggcgccat tctatcctct agaggatgga accgctggag
      241 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
      301 atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc gttcggttgg
      361 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
      421 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
      481 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt tcgcagccta
      541 ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaaattac
      601 caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
      661 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtaccag
      721 agtcctttga tcgtgacaaa acaattgcac tgataatgaa ttcctctgga tctactgggt
      781 tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg catgccagag
      841 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
      901 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
      961 taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac aaaattcaaa
     1021 gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg attgacaaat
     1081 acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg aaagaagtcg
     1141 gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat gggctcactg
     1201 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     1261 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     1321 ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt tatgtaaaca
     1381 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag
     1441 cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct ttaattaaat
     1501 acaaagggta tcaggtggcc cccgctgaat tggaatcgat attgttacaa caccccaaca
     1561 tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg
     1621 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca
     1681 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga
     1741 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga
     1801 agggcggaaa gtccaaattg taatagatcc actagttcta gagcggccgc caccgcggtg
     1861 gaggggtaca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1921 aaaaaaaaaa agtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
     1981 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2041 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2101 aatggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2161 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2221 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2281 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2341 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2401 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2461 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2521 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2581 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2641 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2701 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2761 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2821 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2881 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2941 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3001 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3061 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3121 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3181 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3241 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3301 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3361 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3421 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3481 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3541 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3601 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3661 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3721 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3781 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3841 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3901 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3961 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4021 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4081 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4141 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4201 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4261 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4321 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4381 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4441 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4501 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4561 gtttcccgac tggaaagcgg gcagtgagcg caacgcaat
//
