LOCUS       sequence-243514-        4620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express truncated 3xFLAG tagged firefly luciferase (FFLuc) from AUG 
            at codon 30.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4620)
  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 4620)
  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(30)-FFLuc-3XFLAG
FEATURES             Location/Qualifiers
     source          1..4620
                     /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             2062..2127
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2174..2295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2185..2204)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2239..2258
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2543..2560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2714..3302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2794..2813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3502..4293,4294..4362))
                     /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    4467..4515
                     /note="synthetic polyadenylation signal"
     misc_feature    4529..4620
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4569..4588
                     /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 gaagcgctac gccctggtgc ccggcaccat cgcctttacc
      541 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
      601 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
      661 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
      721 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
      781 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
      841 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
      901 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
      961 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     1021 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     1081 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     1141 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     1201 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     1261 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     1321 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     1381 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     1441 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     1501 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     1561 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     1621 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     1681 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     1741 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     1801 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     1861 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     1921 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     1981 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     2041 aagggcggca agatcgccgt ggactacaaa gaccatgacg gtgattataa agatcatgac
     2101 atcgattaca aggatgacga tgacaagtaa taattctaga gtcggggcgg ccggccgctt
     2161 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2221 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2281 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2341 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     2401 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     2461 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     2521 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2881 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3121 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca
     3481 aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat
     3541 ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg
     3601 tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc
     3661 ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc
     3721 tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc
     3781 gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc
     3841 gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc
     3901 acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa
     3961 gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat
     4021 gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta
     4081 gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca
     4141 tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag
     4201 gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc
     4261 agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc
     4321 aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata
     4381 ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa gtaatattaa
     4441 ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta catctgtgtg
     4501 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     4561 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
