LOCUS       sequence-153928-        6345 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 6345)
  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 6345)
  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-PSS1
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     261..283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     275..292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     276..292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     299..318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        299..317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        810..1021
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1034..1302
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1368..1942,1943..1945,1946..1954)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1535..1552)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1762..1781
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1967..2761
                     /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(2021..2040)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2631..2650
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2821..2841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2883..3004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2920..2939)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2974..2993
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3833..3851)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3872..3888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3872..3888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3885..3907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3896..3912
                     /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(3920..3926,3927..3944,3945..3950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3965..3986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4103..4120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4274..4862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4354..4373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5033..5824,5825..5893))
                     /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     5656..5675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5894..5998)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6024..6345,1..134))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6161..6182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
       61 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
      121 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      181 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
      241 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
      301 atacgactca ctatagggcg aattgggtac cgcgtttaaa ctcgagctgc attccagtgt
      361 tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat
      421 atttacactt acatactaat aataaattca acaaacaatt tatttatgtt tatttattta
      481 ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca
      541 gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc
      601 gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt
      661 gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc
      721 gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaggcctc cacggccaag
      781 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
      841 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
      901 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
      961 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     1021 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     1081 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     1141 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     1201 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     1261 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc
     1321 tactctagag ctagcgaatt cgaatttaaa tcggatccgc ggccgcgccc ctctccctcc
     1381 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1441 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1501 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1561 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1621 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1681 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     1741 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     1801 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     1861 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     1921 ctttgaaaaa cacgatgata atatggccac aaccatggcg tccggaatga ttgaacaaga
     1981 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2041 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2101 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2161 gcggctatcg tggctggccg cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2221 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2281 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2341 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2401 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2461 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     2521 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2581 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2641 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2701 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     2761 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2821 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgcgttaac
     2881 taaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2941 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3001 cttatcatgt ctggaattga ctcaaatgat gtcaattagt ctatcagaag ctcatctggt
     3061 ctcccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt tcccaaactg
     3121 ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac
     3181 gaactcccta aagaaacagt ggcagccagg tttagccccg gaattgactg gattcctttt
     3241 ttagggccca ttggtatggc tttttccccg tatcccccca ggtgtctgca ggctcaaaga
     3301 gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc
     3361 cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc
     3421 ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg
     3481 ggggctgtcc ctgatatcta taacaagaaa atatatatat aataagttat cacgtaagta
     3541 gaacatgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat
     3601 aatcatgcgt cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc
     3661 attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc
     3721 gacggattcg cgctatttag aaagagagag caatatttca agaatgctgg aatgcagact
     3781 agttctagag gccgaggcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     3841 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3901 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3961 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4021 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4081 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4141 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4201 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4681 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5041 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5101 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5161 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5221 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5281 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5341 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5401 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5461 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5521 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5581 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5641 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5701 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5761 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5821 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5881 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6001 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     6061 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6121 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6181 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6241 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6301 aaatcggaac cctaaaggga gcccccgatt tagagcttga cgggg
//
