LOCUS       sequence-116691-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Competitor vector for transcriptional mutagenesis assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  AUTHORS   You C, Wang Y
  TITLE     Quantitative measurement of transcriptional inhibition and 
            mutagenesis induced by site-specifically incorporated DNA lesions in
            vitro and in vivo.
  JOURNAL   Nat Protoc. 2015 Sep;10(9):1389-406. doi: 10.1038/nprot.2015.094. 
            Epub 2015 Aug 20.
  PUBMED    26292071
REFERENCE   2  (bases 1 to 4499)
  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: 
            "10.1038/nprot.2015.094"; journalName: "Nat Protoc"; date: 
            "2015-09"; volume: "10"; issue: "9"; pages: "1389-406"
COMMENT     Sequence Label: pTGFP-T7-Hha10comp
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     585..604
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        585..603
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      708..717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             714..1412
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRINGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     polyA_signal    1535..1656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1572..1591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1626..1645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1663..2118)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2012..2031
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2145..2249
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2274..2294)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             2395..3189
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2395..3189
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2449..2468)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3059..3078
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3377..3396)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3421..3468
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3797..4385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4286..4305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggttta actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtctaatac gactcactat
      601 agggagatcc gctagcggat gcatcgactc aattatagca tatcgccatg gatgactcgc
      661 tgcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggaga
      721 gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaacggc accctgaacg
      781 gcgtggagtt cgagctggtg ggcggcggag agggcacccc cgagcagggc cgcatgacca
      841 acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga
      901 tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc
      961 acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg
     1021 tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg
     1081 tgatgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca
     1141 acgccaccgt ggagcacctg caccccatgg gcgataacga tctggatggc agcttcaccc
     1201 gcaccttcag cctgcgcgac ggcggctact acagctccgt ggtggacagc cacatgcact
     1261 tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc
     1321 gcgtggagga ggatcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca
     1381 agaccccgga tgcagatgcc ggtgaagaat aaagcggccg caactctaga tcataatcag
     1441 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1501 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1561 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1621 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     1681 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1741 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1801 cagtttggaa caaaagtcca ctattaaaga acgtgaactc caacgtcaaa gggcgaaaaa
     1861 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1921 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1981 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2041 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2101 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2221 tgcttcaata atattgaaaa aggaaaagtc ctgaggcgga aagaaccagc tgtggaatgt
     2281 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2341 ggatcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2401 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2461 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2521 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     2581 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2641 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2701 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2761 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2821 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2881 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     2941 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3001 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3061 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3121 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3181 ttcttctgag cggaactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3241 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3301 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3361 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3421 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     3481 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3541 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3601 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     3661 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
