LOCUS       sequence-428796-        8038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SM(FLAG)-optimal Firefly luciferase - 3'UTR malat1 triple 
            helix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8038)
  AUTHORS   Barrington CL, Galindo G, Koch AL, Horton ER, Morrison EJ, Tisa S, 
            Stasevich TJ, Rissland OS
  TITLE     Synonymous codon usage regulates translation initiation.
  JOURNAL   Cell Rep. 2023 Dec 26;42(12):113413. doi: 
            10.1016/j.celrep.2023.113413. Epub 2023 Dec 12.
  PUBMED    38096059
REFERENCE   2  (bases 1 to 8038)
  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: "Cell Rep.";
            date: "2023-12-26"; volume: "42(12):113413. doi"; pages: " 
            10.1016/j.celrep.2023.113413. Epub 2023 Dec 12"
COMMENT     Sequence Label: pcDNA3.1_SM-FLuc100-malat1
FEATURES             Location/Qualifiers
     source          1..8038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(100..204)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(279..298)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        470..849
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        850..1053
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1004..1024
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1098..1117
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1098..1116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1142..1151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1151..1174
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1178..1201
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1205..1228
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1733..1756
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1763..1786
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1799..1822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1829..1852
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2045..2068
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2072..2095
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2099..2122
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      4140..4568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4577..4597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4582..4911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4762..4897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4824..4843
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4978..5772
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5032..5051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5642..5661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5946..6067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5983..6002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6037..6056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6116..6132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6116..6132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6129..6151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6140..6156
                     /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)."
     promoter        complement(join(6164..6170,6171..6188,6189..6194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6209..6230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6347..6364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6518..7106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6598..6617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7277..8038,1..30,31..99))
                     /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     7900..7919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
       61 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
      241 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
      301 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
      361 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
      421 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      481 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      541 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      601 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      661 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      721 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      781 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      841 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      901 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      961 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     1021 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     1081 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgcggc
     1141 cgccaccatg gactacaagg acgacgacga caaaggtgac tacaaagatg atgacgataa
     1201 aggcgactat aaggacgatg acgacaaggg cggaaactca ctgatcaagg aaaacatgcg
     1261 gatgaaggtg gtgatggagg gctccgtgaa tggtcaccag ttcaagtgca ccggagaggg
     1321 agagggaaac ccgtacatgg gaactcagac catgcgcatt aaggtcatcg aaggaggtcc
     1381 gctgccgttc gctttcgata tcctggccac ttcgttcgga ggagggtcgc gcacgttcat
     1441 caagtacccg aagggaatcc cggacttctt taagcagtca ttcccggaag gattcacttg
     1501 ggaacgggtg acccggtatg aagatggagg tgtggtgact gtcatgcaag atacttcgct
     1561 ggaggatggg tgcctcgtgt accacgtcca agtccgcgga gtgaatttcc cgtccaacgg
     1621 accagtgatg cagaaaaaga cgaagggttg ggaacctaat actgaaatga tgtaccccgc
     1681 agacggaggg ctgaggggct acacccacat ggcgctgaag gtcgacggag gagattacaa
     1741 ggatgacgac gataagcaac aagattacaa agacgatgat gacaagggcc agcagggcga
     1801 ctacaaggac gacgacgaca agcagcagga ctacaaagat gacgatgata aaggaggagg
     1861 acatctgtcc tgttcgttcg tgaccaccta cagatcaaag aaaaccgtgg gaaacatcaa
     1921 gatgccgggc attcatgccg tcgaccaccg cctggagcgg ctcgaagaat cagacaatga
     1981 gatgttcgtc gtgcaaagag aacatgccgt ggccaagttc gcgggactgg gaggcggtgg
     2041 aggcgattac aaagacgatg atgacaaggg tgactataaa gacgacgatg acaaagggga
     2101 ttacaaggat gatgatgata agggaggcgg tggatcaggt ggaggaggtt cactgcagga
     2161 ggacgccaaa aacattaaaa aaggcccggc tccattctac ccgctggagg acggtactgc
     2221 tggcgagcag ctgcacaaag ctatgaaacg atacgcacta gttccgggta ctattgcttt
     2281 cactgacgct cacatcgagg tggacatcac ttacgctgag tacttcgaga tgagcgttcg
     2341 gttggccgag gctatgaaac gatacggcct gaacactaac caccgaatcg tggtgtgctc
     2401 agagaactca ttacagttct tcatgccggt gttgggcgcc ttattcatcg gcgttgctgt
     2461 tgccccagcc aacgacattt acaacgagcg agagttgtta aactcaatgg gcatttcaca
     2521 gccaactgtg gtgttcgttt caaaaaaagg cttgcagaaa attttgaacg tgcagaaaaa
     2581 attaccaatc atccagaaaa ttattatcat ggacagcaaa actgactacc agggtttcca
     2641 gagcatgtac actttcgtta cttcacactt gccaccgggt ttcaacgagt acgacttcgt
     2701 gccagagagc ttcgaccgag acaaaactat tgccctgatc atgaactcat caggctcaac
     2761 tggtctgccg aaaggtgttg ctctgccgca ccggactgcc tgcgtgcgat tctcacacgc
     2821 ccgggaccca attttcggca accagatcat tccggacact gccattttaa gcgttgttcc
     2881 attccaccac ggtttcggca tgttcactac tctgggttac ttcatatgcg gtttccgagt
     2941 tgtgttaatg taccggttcg aggaggagct gttcctgcga agcttgcagg actacaaaat
     3001 tcagtcagct ctgctggtgc caactctgtt cagcttcttc gccaaaagca ctctgattga
     3061 caaatacgac ttatcaaact tacacgagat tgcttcaggt ggcgctccgt tatcaaaaga
     3121 ggttggcgag gccgttgcca aacgattcca cctgccaggt atccgacagg gttacggttt
     3181 gactgagact acttcagcta ttctgattac tccggagggt gacgacaaac cgggcgctgt
     3241 gggtaaagtt gttccattct tcgaggctaa agttgtggac ctggacactg gtaaaactct
     3301 gggcgttaac cagcgaggcg agctgtgcgt gcgaggtccg atgattatga gcggttacgt
     3361 gaacaacccg gaggctacta acgccttgat tgacaaagac ggttggttac acagcggtga
     3421 catcgcttac tgggacgagg acgagcactt cttcatcgtt gaccggctga aatcactgat
     3481 taaatacaaa ggctaccagg tggctccggc tgagttggag tcaatcttgt tacagcaccc
     3541 aaacatcttc gacgctggtg tggctggtct gccggacgac gacgccggtg agttaccagc
     3601 cgccgttgtt gttttggagc acggtaaaac tatgactgag aaagagatcg tggactacgt
     3661 tgccagccag gttactactg ctaaaaaatt gcggggtggc gttgtgttcg tcgacgaggt
     3721 tccgaaaggt ttaactggca aattggacgc tcgaaaaatc cgagagatcc tgatcaaagc
     3781 caaaaaaggc ggcaaaatcg ccgtgtaagg taccgagctc ggatccacta gtccagtgtg
     3841 gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     3901 acccgctgat cagcctcgac gattcgtcag tagggttgta aaggtttttc ttttcctgag
     3961 aaaacaacct tttgttttct caggttttgc tttttggcct ttccctagct ttaaaaaaaa
     4021 aaaagcaaaa gacgctggtg gctggcactc ctggtttcca ggacggggtt caagtccctg
     4081 cggtgtcttt gcttcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4681 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4741 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4801 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4861 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4921 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4981 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5041 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5101 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5161 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5221 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5281 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5341 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5401 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5461 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5521 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5581 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5641 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5701 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5761 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5821 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5881 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5941 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6061 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6301 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6361 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6421 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6481 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6541 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6601 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6661 ttaccggata cctgtccgcc tttttccctt cgggaagcgt ggcgctttct catagctcac
     6721 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6781 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6841 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6901 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6961 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7021 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7321 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7381 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7441 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7501 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7561 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7621 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7681 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7741 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7801 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7861 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7921 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7981 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagc
//
