LOCUS       sequence-116688-        4496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Parent vector for constructing non-replicating lesion-bearing 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4496)
  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 4496)
  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-Hha10T
FEATURES             Location/Qualifiers
     source          1..4496
                     /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      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             711..1409
                     /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    1532..1653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1569..1588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1623..1642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1660..2115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2009..2028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2142..2246
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2271..2291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             2392..3186
                     /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             2392..3186
                     /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(2446..2465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3056..3075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3374..3393)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3418..3465
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3794..4382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4283..4302
                     /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 cgccatggat gactcgctgc
      661 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggagagcg
      721 acgagagcgg cctgcccgcc atggagatcg agtgccgcat caacggcacc ctgaacggcg
      781 tggagttcga gctggtgggc ggcggagagg gcacccccga gcagggccgc atgaccaaca
      841 agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg
      901 gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg
      961 ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc
     1021 tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtga
     1081 tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg
     1141 ccaccgtgga gcacctgcac cccatgggcg ataacgatct ggatggcagc ttcacccgca
     1201 ccttcagcct gcgcgacggc ggctactaca gctccgtggt ggacagccac atgcacttca
     1261 agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg
     1321 tggaggagga tcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga
     1381 ccccggatgc agatgccggt gaagaataaa gcggccgcaa ctctagatca taatcagcca
     1441 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1501 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     1561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1621 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     1681 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1741 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     1801 tttggaacaa aagtccacta ttaaagaacg tgaactccaa cgtcaaaggg cgaaaaaccg
     1861 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     1921 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     1981 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     2041 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2101 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2161 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2221 ttcaataata ttgaaaaagg aaaagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     2281 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgga
     2341 tcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     2401 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2461 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2521 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     2581 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2641 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2701 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2761 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     2821 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     2881 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     2941 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3001 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3061 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3121 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3181 ttctgagcgg aactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3241 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3301 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     3361 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3421 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     3481 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     3541 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     3601 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     3661 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3721 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3781 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3841 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3901 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3961 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4021 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4081 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4141 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4201 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4261 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4321 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4381 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4441 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
