LOCUS       sequence-152226-        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     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        814..1025
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1038..1306
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1372..1946,1947..1949,1950..1958)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1539..1556)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1766..1785
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1971..2765
                     /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(2025..2044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2635..2654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2825..2845)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2887..3008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2924..2943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2978..2997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3841..3859)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3880..3896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3880..3896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3893..3915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3904..3920
                     /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(3928..3934,3935..3952,3953..3958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3973..3994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4111..4128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4282..4870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4362..4381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5041..5832,5833..5901))
                     /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     5664..5683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5902..6006)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6032..6353,1..134))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6169..6190)
                     /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 ctcgagttaa ccctagaaag
      361 atgttttatc ggtctgtata tcgaggttta tttattaatt tgaatagata ttaagtttta
      421 ttatatttac acttacatac taataataaa ttcaacaaac aatttattta tgtttattta
      481 tttattaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttatgagg
      541 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
      601 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
      661 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
      721 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaagg cctccacggc
      781 caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
      841 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
      901 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
      961 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     1021 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     1081 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     1141 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     1201 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     1261 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     1321 cgcctactct agagctagcg aattcgaatt taaatcggat ccgcggccgc gcccctctcc
     1381 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     1441 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     1501 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     1561 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     1621 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     1681 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1741 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1801 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     1861 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     1921 tttcctttga aaaacacgat gataatatgg ccacaaccat ggcgtccgga atgattgaac
     1981 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2041 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2101 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2161 cagcgcggct atcgtggctg gccgcgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2221 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2281 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2341 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2401 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2461 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2521 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2581 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2641 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2701 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2761 tctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2821 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcgt
     2881 taactaaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2941 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3001 gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat tagtctatca gaagctcatc
     3061 tggtctccct tccgggggac aagacatccc tgtttaatat ttaaacagca gtgttcccaa
     3121 actgggttct tatatccctt gctctggtca accaggttgc agggtttcct gtcctcacag
     3181 gaacgaagtc cctaaagaaa cagtggcagc caggtttagc cccggaattg actggattcc
     3241 ttttttaggg cccattggta tggctttttc cccgtatccc cccaggtgtc tgcaggctca
     3301 aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc
     3361 tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc
     3421 gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg
     3481 ggggggggct gtccctgata tctataacaa gaaaatatat atataataag ttatcacgta
     3541 agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa
     3601 agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta
     3661 ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca
     3721 cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg tctttctagg
     3781 gttaaactag ttctagaggc cgaggcggcc gccaccgcgg tggagctcca gcttttgttc
     3841 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4681 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     4741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6001 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     6061 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6121 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6181 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6241 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6301 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg ggg
//
