LOCUS       sequence-152210-        6323 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 6323)
  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 6323)
  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-Control
FEATURES             Location/Qualifiers
     source          1..6323
                     /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        799..1010
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1023..1291
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    join(1357..1931,1932..1934,1935..1943)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1524..1541)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1751..1770
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1956..2750
                     /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(2010..2029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2620..2639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2810..2830)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2872..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2909..2928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2963..2982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(3811..3829)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3850..3866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3850..3866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3863..3885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3874..3890
                     /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(3898..3904,3905..3922,3923..3928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3943..3964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4081..4098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4252..4840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4332..4351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5011..5802,5803..5871))
                     /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     5634..5653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5872..5976)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6002..6323,1..134))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6139..6160)
                     /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 ctcgagtgtt ttatcggtct
      361 gtatatcgag gtttatttat taatttgaat agatattaag ttttattata tttacactta
      421 catactaata ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac
      481 aaaaactcaa aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca
      541 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
      601 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
      661 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
      721 agcctgcaga cacctggggg gatacgggga aaaggcctcc acggccaagg atctgcgatc
      781 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
      841 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      901 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      961 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     1021 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     1081 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     1141 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     1201 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     1261 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct actctagagc
     1321 tagcgaattc gaatttaaat cggatccgcg gccgcgcccc tctccctccc ccccccctaa
     1381 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1441 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1501 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1561 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1621 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     1681 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     1741 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     1801 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     1861 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     1921 acgatgataa tatggccaca accatggcgt ccggaatgat tgaacaagat ggattgcacg
     1981 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2041 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2101 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2161 ggctggccgc gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2221 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2281 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2341 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2401 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2461 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2521 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2581 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2641 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2701 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gtcgacaatc
     2761 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2821 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgcgttaact aaacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3001 tggaattgac tcaaatgatg tcaattagtc tatcagaagc tcatctggtc tcccttccgg
     3061 gggacaagac atccctgttt aatatttaaa cagcagtgtt cccaaactgg gttcttatat
     3121 cccttgctct ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg aagtccctaa
     3181 agaaacagtg gcagccaggt ttagccccgg aattgactgg attccttttt tagggcccat
     3241 tggtatggct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag
     3301 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg
     3361 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct
     3421 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc
     3481 tgatatctat aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat
     3541 aacaatataa ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc
     3601 attttgactc acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca
     3661 cgcctcacgg gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc
     3721 gctatttaga aagagagagc aatatttcaa gaatgactag ttctagaggc cgaggcggcc
     3781 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc
     3841 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3901 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3961 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4021 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4081 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4141 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4201 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4681 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5041 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5101 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5161 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5221 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5281 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5341 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5401 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5461 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5521 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5581 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5641 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5701 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5761 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5821 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5881 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5941 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6001 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     6061 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     6121 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     6181 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     6241 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     6301 ccccgattta gagcttgacg ggg
//
