LOCUS       sequence-367120-        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     73..95
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     87..104
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     88..104
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..130
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        622..833
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             846..1114
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1180..1754,1755..1757,1758..1766)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1347..1364)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1574..1593
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1779..2573
                     /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(1833..1852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2443..2462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2633..2653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2695..2816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2732..2751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2786..2805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3645..3663)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3684..3700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3684..3700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3697..3719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3708..3724
                     /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(3732..3738,3739..3756,3757..3762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3777..3798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3915..3932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4086..4674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4166..4185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4845..5636,5637..5705))
                     /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     5468..5487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5706..5810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5836..6291)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5973..5994)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6185..6204
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
       61 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
      121 cactataggg cgaattgggt accgcgttta aactcgagct gcattccagt gttttatcgg
      181 tctgtatatc gaggtttatt tattaatttg aatagatatt aagttttatt atatttacac
      241 ttacatacta ataataaatt caacaaacaa tttatttatg tttatttatt tattaaaaaa
      301 aacaaaaact caaaatttct tctataaagt aacaaaactt ttatgaggga cagccccccc
      361 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
      421 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
      481 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
      541 ttgagcctgc agacacctgg ggggatacgg ggaaaaggcc tccacggcca aggatctgcg
      601 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      661 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
      721 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      781 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
      841 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
      901 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
      961 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     1021 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     1081 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctactctag
     1141 agctagcgaa ttcgaattta aatcggatcc gcggccgcgc ccctctccct cccccccccc
     1201 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1261 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     1321 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     1381 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     1441 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     1501 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     1561 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     1621 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     1681 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     1741 aacacgatga taatatggcc acaaccatgg cgtccggaat gattgaacaa gatggattgc
     1801 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1861 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1921 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     1981 cgtggctggc cgcgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2041 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2101 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2161 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2221 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2281 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2341 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2401 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2461 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2521 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtcgaca
     2581 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2641 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgcgtta actaaacttg
     2701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2821 gtctggaatt gactcaaatg atgtcaatta gtctatcaga agctcatctg gtctcccttc
     2881 cgggggacaa gacatccctg tttaatattt aaacagcagt gttcccaaac tgggttctta
     2941 tatcccttgc tctggtcaac caggttgcag ggtttcctgt cctcacagga acgaagtccc
     3001 taaagaaaca gtggcagcca ggtttagccc cggaattgac tggattcctt ttttagggcc
     3061 cattggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     3121 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     3181 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     3241 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     3301 ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
     3361 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
     3421 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
     3481 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
     3541 cgcgctattt agaaagagag agcaatattt caagaatgct ggaatgcaga ctagttctag
     3601 aggccgaggc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     3661 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3721 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3781 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3841 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3901 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3961 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4021 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4501 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4561 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4621 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4681 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4741 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4801 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4861 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4921 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4981 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5041 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5101 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5161 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5221 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5281 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5341 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5401 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5461 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5521 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5581 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5641 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5701 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5761 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5821 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5881 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5941 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     6001 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     6061 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     6121 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     6181 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     6241 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     6301 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcct
//
