LOCUS       sequence-243515-        4533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express truncated 3xFLAG tagged firefly luciferase (FFLuc) from AUG 
            at codon 59.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4533)
  AUTHORS   Kearse MG, Goldman DH, Choi J, Nwaezeapu C, Liang D, Green KM, 
            Goldstrohm AC, Todd PK, Green R, Wilusz JE
  TITLE     Ribosome queuing enables non-AUG translation to be resistant to 
            multiple protein synthesis inhibitors.
  JOURNAL   Genes Dev. 2019 Jun 6. pii: gad.324715.119. doi: 
            10.1101/gad.324715.119.
  PUBMED    31171704
REFERENCE   2  (bases 1 to 4533)
  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: "Genes Dev. 
            2019 Jun 6. pii: gad.324715.119. doi: 10.1101/gad.324715.119."
COMMENT     Sequence Label: pGL4.13/AUG(59)-FFLuc-3XFLAG
FEATURES             Location/Qualifiers
     source          1..4533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        106..297
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(129..149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      314..449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1975..2040
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2087..2208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2098..2117)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2152..2171
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2456..2473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2627..3215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2707..2726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3415..4206,4207..4275))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4380..4428
                     /note="synthetic polyadenylation signal"
     misc_feature    4442..4533
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4482..4501
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcagatct gcgcagcacc
       61 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
      121 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
      181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
      241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
      301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
      361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
      421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg gcaatccggt
      481 actgttggta aagccaccat gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg
      541 aatacaaacc atcggatcgt ggtgtgcagc gagaatagct tgcagttctt catgcccgtg
      601 ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta acgacatcta caacgagcgc
      661 gagctgctga acagcatggg catcagccag cccaccgtcg tattcgtgag caagaaaggg
      721 ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat catcatcatg
      781 gatagcaaga ccgactacca gggcttccaa agcatgtaca ccttcgtgac ttcccatttg
      841 ccacccggct tcaacgagta cgacttcgtg cccgagagct tcgaccggga caaaaccatc
      901 gccctgatca tgaacagtag tggcagtacc ggattgccca agggcgtagc cctaccgcac
      961 cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc
     1021 cccgacaccg ctatcctcag cgtggtgcca tttcaccacg gcttcggcat gttcaccacg
     1081 ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga ggaggagcta
     1141 ttcttgcgca gcttgcaaga ctataagatt caatctgccc tgctggtgcc cacactattt
     1201 agcttcttcg ctaagagcac tctcatcgac aagtacgacc taagcaactt gcacgagatc
     1261 gccagcggcg gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa acgcttccac
     1321 ctaccaggca tccgccaggg ctacggcctg acagaaacaa ccagcgccat tctgatcacc
     1381 cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag
     1441 gtggtggact tggacaccgg taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc
     1501 cgtggcccca tgatcatgag cggctacgtt aacaaccccg aggctacaaa cgctctcatc
     1561 gacaaggacg gctggctgca cagcggcgac atcgcctact gggacgagga cgagcacttc
     1621 ttcatcgtgg accggctgaa gagcctgatc aaatacaagg gctaccaggt agccccagcc
     1681 gaactggaga gcatcctgct gcaacacccc aacatcttcg acgccggggt cgccggcctg
     1741 cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc
     1801 atgaccgaga aggagatcgt ggactatgtg gccagccagg ttacaaccgc caagaagctg
     1861 cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc
     1921 cgcaagatcc gcgagattct cattaaggcc aagaagggcg gcaagatcgc cgtggactac
     1981 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     2041 taataattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     2101 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2161 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2221 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2281 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     2341 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2401 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2821 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3061 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3361 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     3421 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     3481 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     3541 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     3601 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     3661 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     3721 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     3781 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     3841 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     3901 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     3961 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     4021 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     4081 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     4141 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     4201 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     4261 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     4321 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
     4381 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     4441 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     4501 ccagtgcaag tgcaggtgcc agaacatttc tct
//
