LOCUS       sequence-267634-        7035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector for constitutive expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7035)
  AUTHORS   Fischer AD, Veronese-Paniagua DA, Swaminathan S, Kashima H, Rubin 
            DC, Madison BB
  TITLE     The oncogenic function of PLAGL2 is mediated via ASCL2 and IGF2 and 
            a Wnt-independent mechanism in colorectal cancer.
  JOURNAL   Am J Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022.
  PUBMED    37310750
REFERENCE   2  (bases 1 to 7035)
  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: "Am J 
            Physiol Gastrointest Liver Physiol. 2023 Jun 13. doi: 
            10.1152/ajpgi.00058.2022."
COMMENT     Sequence Label: BII-ChPtW
FEATURES             Location/Qualifiers
     source          1..7035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   532..566
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(835..851)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(835..851)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2402..2455
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2462..2488
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2462..2488
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2538..3126
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2591..2611)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3009..3020)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3206..3340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3230..3249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3284..3303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4119..4181
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4311..4328)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4311..4327)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4320..4342)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4469..4924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4556..4575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4766..4787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4950..5054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5055..5123,5124..5915)
                     /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(5273..5292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6086..6674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6575..6594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6828..6845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6962..6983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6998..7003,7004..7021,7022..7028)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
       61 cccgccgggt aactcacggg gtatccatgt ccatttctgc ggcatccagc caggataccc
      121 gtcctcgctg acgtaatatc ccagcgccgc accgctgtca ttaatctgca caccggcacg
      181 gcagttccgg ctgtcgccgg tattgttcgg gttgctgatg cgcttcgggc tgaccatccg
      241 gaactgtgtc cggaaaagcc gcgacgaact ggtatcccag gtggcctgaa cgaacagttc
      301 accgttaaag gcgtgcatgg ccacaccttc ccgaatcatc atggtaaacg tgcgttttcg
      361 ctcaacgtca atgcagcagc agtcatcctc ggcaaactct ttccatgccg cttcaacctc
      421 gcgggaaaag gcacgggctt cttcctcccc gatgcccaga tagcgccagc ttgggcgatg
      481 actgagccgg aaaaaagacc cgacgatatg atcctgatgc agctagatta accctagaaa
      541 gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct ttctaaatag
      601 cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct cccgtgaggc
      661 gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg tgagtcaaaa
      721 tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata attatattgt
      781 tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt tatagatcca
      841 ctagttctag agggacagcc cccccccaaa gcccccaggg atgtaattac gtccctcccc
      901 cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat
      961 ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt
     1021 tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa
     1081 aggcctccaa ggcctgcagg agtcaatggg aaaaacccat tggagccaag tacactgact
     1141 caatagggac tttccattgg gttttgccca gtacataagg tcaatagggg gtgagtcaac
     1201 aggaaagtcc cattggagcc aagtacattg agtcaatagg gactttccaa tgggttttgc
     1261 ccagtacata aggtcaatgg gaggtaagcc aatgggtttt tcccattact gacatgtata
     1321 ctgagtcatt agggactttc caatgggttt tgcccagtac ataaggtcaa taggggtgaa
     1381 tcaacaggaa agtcccattg gagccaagta cactgagtca atagggactt tccattgggt
     1441 tttgcccagt acaaaaggtc aatagggggt gagtcaatgg gtttttccca ttattggcac
     1501 atacataagg tcaatagggg tgactagtgg agaagagcat gcttgagggc tgagtgcccc
     1561 tcagtgggca gagagcacat ggcccacagt ccctgagaag ttggggggag gggtgggcaa
     1621 ttgaactggt gcctagagaa ggtggggctt gggtaaactg ggaaagtgat gtggtgtact
     1681 ggctccacct ttttccccag ggtgggggag aaccatatat aagtgcagta gtctctgtga
     1741 acattcggtt tagtgaacct gcaggtcggc cgccacgacc ggtgccacca tgactgaata
     1801 caaaccaact gttcgcctgg caactcgtga tgatgttcca cgtgcagttc gcaccctggc
     1861 tgctgcattt gctgactacc ctgcaacccg tcacactgtg gacccagacc gccacattga
     1921 acgtgtgact gaactgcagg agctgttcct gacccgtgtg ggcctggaca ttggcaaagt
     1981 gtgggtggca gatgatggtg ctgctgtggc agtgtggacc acccctgaat ctgttgaagc
     2041 tggtgcagtg tttgctgaga ttggcccacg catggcagaa ctgtctggca gccgcctggc
     2101 agcacaacag cagatggaag gtctgctggc accacaccgc ccaaaagaac ctgcttggtt
     2161 cctggcaact gtgggtgtga gccctgacca ccagggtaag ggcctgggct ctgcagtggt
     2221 gctgcctggt gtggaagcag ctgaacgtgc aggtgtgcct gctttcctgg agacctcagc
     2281 tccacgcaac ctgcctttct atgaacgcct gggcttcact gtgactgctg atgtggaagt
     2341 gccagaaggc ccacgcactt ggtgcatgac tcgcaaacca ggtgctagcc ttggctccgg
     2401 cgagggcagg ggaagtcttc taacatgcgg ggacgtggag gaaaatcccg gcccactcgg
     2461 ctacccctac gacgtgcccg actacgccag cggcatgaga cggatagctc gtctcagcta
     2521 gagttcgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2581 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2641 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2701 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2761 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2821 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2881 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     2941 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3001 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3061 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3121 ccccgcctgg aattaattct ggctaagcac ttcgtggccg tcgatcgttt aaagggaggt
     3181 agtgagtcga gatatcgata ctagtgatcc agacatgata agatacattg atgagtttgg
     3241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3301 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3361 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3421 caaatgtggt atggctgatt atgatcctct agagggacag ccccccccca aagcccccag
     3481 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     3541 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     3601 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     3661 cacctggggg gatacgggga aaaagccata ccaatggagc tcacggggac agcccccccc
     3721 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
     3781 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
     3841 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
     3901 tgagcctgca gacacctggg gggatacggg gaaaaagaat tcgataaaag ttttgttact
     3961 ttatagaaga aattttgagt ttttgttttt tttaataaat aaataaacat aaataaattg
     4021 tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa
     4081 attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacacatga
     4141 ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaatctag ctgcgtgttc
     4201 tgcagcgtgt cgagcatctt catctgctcc atcacgctgt aaaacacatt tgcaccgcga
     4261 gtctgcccgt cctccacggg ttcaaaaacg tgaatgaacg aggcgcgctc actggccgtc
     4321 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4381 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4441 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4921 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5041 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5101 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5161 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5221 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5281 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5341 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5401 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5461 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5521 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5581 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5641 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5701 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5761 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5821 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5881 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5941 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6001 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6061 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6841 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6901 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6961 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7021 gtatgttgtg tggaa
//
