LOCUS       sequence-9781-ex        7912 bp    DNA     circular SYN 14-APR-2025
DEFINITION  piggyBac empty vector with Gatweway cassette and CAG constitutive 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7912)
  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 7912)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..7912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2340..2374
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2753..3132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3134..3411
                     /label=chicken beta-actin promoter
     intron          3412..4427
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4351..4373
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4435..4454
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    4521..4645
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4670..4675,4676..4693,4694..4700)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4754..5413
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4820..4839)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5755..6060
                     /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     5939..5958
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6101..6225)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6321..6340)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6386..6441
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6440..6459)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   7145..7207
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7273..7292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7274..7292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7299..7316)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7299..7315)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7308..7330)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7457..7885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7544..7563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7754..7775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg
     2221 gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca
     2281 aacgaagatt ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac
     2341 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
     2401 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
     2461 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
     2521 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
     2581 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
     2641 tagatatcgt gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc
     2701 tctgctcttc tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tcgacattga
     2761 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2821 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2881 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2941 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     3001 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3061 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     3121 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     3181 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     3241 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
     3301 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     3361 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     3421 gcgcgctgcc ttcgccccgt gccccgctcc gcgccgcctc gcgccgcccg ccccggctct
     3481 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
     3541 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
     3601 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
     3661 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
     3721 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     3781 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     3841 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     3901 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     3961 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     4021 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     4081 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     4141 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     4201 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     4261 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     4321 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     4381 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     4441 tgctggttat tgtgctgtct catcattttg gcaaagaatt cgcggccgcg gtaccggcgc
     4501 gccggatcct gcaggttatc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
     4561 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     4621 aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta
     4681 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc
     4741 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     4801 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     4861 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     4921 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     4981 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     5041 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     5101 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     5161 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     5221 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     5281 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     5341 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     5401 gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt atgcgtattt
     5461 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     5521 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     5581 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     5641 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     5701 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     5761 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     5821 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     5881 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     5941 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     6001 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     6061 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     6121 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     6181 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgataa ctctagagaa
     6241 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     6301 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     6361 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     6421 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     6481 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     6541 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     6601 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     6661 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     6721 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     6781 cccaagcttg gatccctcga gttaattaac gagagcataa tattgatatg tgccaaagtt
     6841 gtttctgact gactaataag tataatttgt ttctattatg tataggttaa gctaattact
     6901 tattttataa tacaacatga ctgtttttaa agtacaaaat aagtttattt ttgtaaaaga
     6961 gagaatgttt aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttta
     7021 ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa
     7081 tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac agaccgataa
     7141 aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata tgattatctt
     7201 tctagggtta aataatagtt tctaattttt ttattattca gcctgctgtc gtgaataccg
     7261 agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     7321 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7381 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7441 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7861 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat tt
//
