LOCUS       sequence-366184-        6353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon Reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6353)
  AUTHORS   Henssen AG, Koche R, Zhuang J, Jiang E, Reed C, Eisenberg A, Still 
            E, MacArthur IC, Rodriguez-Fos E, Gonzalez S, Puiggros M, Blackford 
            AN, Mason CE, de Stanchina E, Gonen M, Emde AK, Shah M, Arora K, 
            Reeves C, Socci ND, Perlman E, Antonescu CR, Roberts CWM, Steen H, 
            Mullen E, Jackson SP, Torrents D, Weng Z, Armstrong SA, Kentsis A
  TITLE     PGBD5 promotes site-specific oncogenic mutations in human tumors.
  JOURNAL   Nat Genet. 2017 May 15. doi: 10.1038/ng.3866.
  PUBMED    28504702
REFERENCE   2  (bases 1 to 6353)
  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: "Nat Genet. 
            2017 May 15. doi: 10.1038/ng.3866."
COMMENT     Sequence Label: pBS-EF1-IRES-NEO-PB-ITR
FEATURES             Location/Qualifiers
     source          1..6353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..94
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     86..103
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     87..103
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     110..129
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        110..128
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        625..836
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             849..1117
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1183..1757,1758..1760,1761..1769)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1350..1367)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1577..1596
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1782..2576
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1836..1855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2446..2465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2636..2656)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2698..2819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2735..2754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2789..2808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3652..3670)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3691..3707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3691..3707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3704..3726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3715..3731
                     /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(3739..3745,3746..3763,3764..3769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3784..3805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3922..3939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4093..4681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4173..4192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4852..5643,5644..5712))
                     /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     5475..5494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5713..5817)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5843..6298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5980..6001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6192..6211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
       61 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
      121 actatagggc gaattgggta ccgcgtttaa actcgagtta accctagaaa gatgttttat
      181 cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt attatattta
      241 cacttacata ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa
      301 aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa cttttatgag ggacagcccc
      361 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
      421 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
      481 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
      541 tctttgagcc tgcagacacc tggggggata cggggaaaag gcctccacgg ccaaggatct
      601 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      661 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
      721 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      781 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
      841 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
      901 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
      961 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     1021 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     1081 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctactc
     1141 tagagctagc gaattcgaat ttaaatcgga tccgcggccg cgcccctctc cctccccccc
     1201 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     1261 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     1321 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     1381 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     1441 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     1501 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     1561 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     1621 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     1681 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     1741 aaaaacacga tgataatatg gccacaacca tggcgtccgg aatgattgaa caagatggat
     1801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     1921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     1981 tatcgtggct ggccgcgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     2041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     2101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     2161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     2221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     2281 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     2341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     2401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     2461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     2521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg
     2581 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2641 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgcg ttaactaaac
     2701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2821 catgtctgga attgactcaa atgatgtcaa ttagtctatc agaagctcat ctggtctccc
     2881 ttccggggga caagacatcc ctgtttaata tttaaacagc agtgttccca aactgggttc
     2941 ttatatccct tgctctggtc aaccaggttg cagggtttcc tgtcctcaca ggaacgaagt
     3001 ccctaaagaa acagtggcag ccaggtttag ccccggaatt gactggattc cttttttagg
     3061 gcccattggt atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
     3121 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc
     3181 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc
     3241 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc
     3301 tgtccctgat atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     3361 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     3421 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     3481 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     3541 attcgcgcta tttagaaaga gagagcaata tttcaagaat gtctttctag ggttaaacta
     3601 gttctagagg ccgaggcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     3661 agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5821 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5881 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5941 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     6001 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     6061 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     6121 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6181 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6241 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     6301 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctc
//
