LOCUS       sequence-174155-        7139 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  piggyBac empty vector with Gatweway cassette and CAG constitutive 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7139)
  AUTHORS   Woltjen K, Michael IP, Mohseni P, Desai R, Mileikovsky M, Hamalainen
            R, Cowling R, Wang W, Liu P, Gertsenstein M, Kaji K, Sung HK, Nagy A
  TITLE     piggyBac transposition reprograms fibroblasts to induced pluripotent
            stem cells.
  JOURNAL   Nature. 2009 Apr 9;458(7239):766-70. doi: 10.1038/nature07863
  PUBMED    19252478
REFERENCE   2  (bases 1 to 7139)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2009-04-9- 9"; volume: "458"; issue: "7239"; pages:
            "766-70"
COMMENT     Sequence Label: PB-CA
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     329..351
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     413..432
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    499..623
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(648..653,654..671,672..678)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             732..1391
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(798..817)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1733..2038
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1917..1936
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2079..2203)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2299..2318)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2364..2419
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2418..2437)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3123..3185
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3251..3270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3252..3270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3277..3294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3277..3293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3286..3308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3435..3890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3522..3541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3732..3753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3916..4020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4021..4089,4090..4881)
                     /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     complement(4239..4258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5052..5640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5541..5560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5794..5811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5928..5949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5964..5969,5970..5987,5988..5994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6002..6018
                     /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)."
     primer_bind     6007..6029
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6026..6042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6026..6042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6063..6081
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   6230..6264
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        6643..7022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
       61 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
      121 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
      181 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
      241 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
      301 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
      361 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
      421 ctggttattg tgctgtctca tcattttggc aaagaattcg cggccgcggt accggcgcgc
      481 cggatcctgc aggttatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
      541 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
      601 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
      661 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
      721 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      781 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
      841 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
      901 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
      961 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1021 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1081 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1141 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1201 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1261 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     1321 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1381 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     1441 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     1501 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     1561 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     1621 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     1681 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     1741 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1801 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1861 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     1921 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     1981 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     2041 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     2101 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     2161 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgataact ctagagaatt
     2221 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     2281 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     2341 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2401 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     2461 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     2521 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     2581 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     2641 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     2701 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     2761 caagcttgga tccctcgagt taattaacga gagcataata ttgatatgtg ccaaagttgt
     2821 ttctgactga ctaataagta taatttgttt ctattatgta taggttaagc taattactta
     2881 ttttataata caacatgact gtttttaaag tacaaaataa gtttattttt gtaaaagaga
     2941 gaatgtttaa aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat
     3001 aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata
     3061 taataaaact taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa
     3121 cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc
     3181 tagggttaaa taatagtttc taattttttt attattcagc ctgctgtcgt gaataccgag
     3241 ctccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     3301 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     3361 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3421 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3481 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3541 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3601 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3661 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3721 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3781 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3841 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     3901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3961 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4021 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4081 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4141 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4201 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4261 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4321 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4381 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4441 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4501 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4561 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4621 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4681 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4741 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4801 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4861 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5761 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5821 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5881 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5941 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6001 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     6061 gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg gtttgccatt
     6121 ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca aacgaagatt
     6181 ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac cctagaaaga
     6241 tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt ctaaatagcg
     6301 cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc cgtgaggcgt
     6361 gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg agtcaaaatg
     6421 acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat tatattgtta
     6481 tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta tagatatcgt
     6541 gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc tctgctcttc
     6601 tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tcgacattga ttattgacta
     6661 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     6721 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     6781 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     6841 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     6901 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     6961 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7021 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     7081 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg ggggggggg
//
