LOCUS       sequence-256367-        7034 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 7034)
  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 7034)
  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..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(308..324)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(308..324)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1875..1928
                     /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             1935..1961
                     /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             1935..1961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2011..2599
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2064..2084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2482..2493)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2679..2813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2757..2776
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3591..3653
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3783..3800)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3783..3799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3792..3814)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3941..4396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4422..4526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4527..4595,4596..5387)
                     /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(4745..4764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5558..6146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6047..6066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6300..6317
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6434..6455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6470..6475,6476..6493,6494..6500)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6508..6524
                     /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     6513..6535
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6532..6548
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6532..6548
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat ccactagttc tagagggaca gccccccccc aaagccccca gggatgtaat
      361 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
      421 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
      481 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
      541 ggatacgggg aaaaggcctc caaggcctgc aggagtcaat gggaaaaacc cattggagcc
      601 aagtacactg actcaatagg gactttccat tgggttttgc ccagtacata aggtcaatag
      661 ggggtgagtc aacaggaaag tcccattgga gccaagtaca ttgagtcaat agggactttc
      721 caatgggttt tgcccagtac ataaggtcaa tgggaggtaa gccaatgggt ttttcccatt
      781 actgacatgt atactgagtc attagggact ttccaatggg ttttgcccag tacataaggt
      841 caataggggt gaatcaacag gaaagtccca ttggagccaa gtacactgag tcaataggga
      901 ctttccattg ggttttgccc agtacaaaag gtcaataggg ggtgagtcaa tgggtttttc
      961 ccattattgg cacatacata aggtcaatag gggtgactag tggagaagag catgcttgag
     1021 ggctgagtgc ccctcagtgg gcagagagca catggcccac agtccctgag aagttggggg
     1081 gaggggtggg caattgaact ggtgcctaga gaaggtgggg cttgggtaaa ctgggaaagt
     1141 gatgtggtgt actggctcca cctttttccc cagggtgggg gagaaccata tataagtgca
     1201 gtagtctctg tgaacattcg gtttagtgaa cctgcaggtc ggccgccacg accggtgcca
     1261 ccatgactga atacaaacca actgttcgcc tggcaactcg tgatgatgtt ccacgtgcag
     1321 ttcgcaccct ggctgctgca tttgctgact accctgcaac ccgtcacact gtggacccag
     1381 accgccacat tgaacgtgtg actgaactgc aggagctgtt cctgacccgt gtgggcctgg
     1441 acattggcaa agtgtgggtg gcagatgatg gtgctgctgt ggcagtgtgg accacccctg
     1501 aatctgttga agctggtgca gtgtttgctg agattggccc acgcatggca gaactgtctg
     1561 gcagccgcct ggcagcacaa cagcagatgg aaggtctgct ggcaccacac cgcccaaaag
     1621 aacctgcttg gttcctggca actgtgggtg tgagccctga ccaccagggt aagggcctgg
     1681 gctctgcagt ggtgctgcct ggtgtggaag cagctgaacg tgcaggtgtg cctgctttcc
     1741 tggagacctc agctccacgc aacctgcctt tctatgaacg cctgggcttc actgtgactg
     1801 ctgatgtgga agtgccagaa ggcccacgca cttggtgcat gactcgcaaa ccaggtgcta
     1861 gccttggctc cggcgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     1921 ccggcccact cggctacccc tacgacgtgc ccgactacgc cagcggcatg agacggatag
     1981 ctcgtctcag ctagagttcg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
     2041 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2101 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2161 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2221 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2281 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2341 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2401 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2461 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2521 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2581 ctttgggccg cctccccgcc tggaattaat tctggctaag cacttcgtgg ccgtcgatcg
     2641 tttaaaggga ggtagtgagt cgagatatcg atactagtga tccagacatg ataagataca
     2701 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2761 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2821 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2881 agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagaggga cagccccccc
     2941 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
     3001 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
     3061 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
     3121 ttgagcctgc agacacctgg ggggatacgg ggaaaagcca taccaatgga gctcacgggg
     3181 acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc
     3241 agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag
     3301 cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt
     3361 ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaaga attcgataaa
     3421 agttttgtta ctttatagaa gaaattttga gtttttgttt tttttaataa ataaataaac
     3481 ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata ataaaactta
     3541 atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca catgcgtcaa
     3601 ttttacacat gattatcttt aacgtacgtc acaatatgat tatctttcta gggttaatct
     3661 agctgcgtgt tctgcagcgt gtcgagcatc ttcatctgct ccatcacgct gtaaaacaca
     3721 tttgcaccgc gagtctgccc gtcctccacg ggttcaaaaa cgtgaatgaa cgaggcgcgc
     3781 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3841 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3901 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4321 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4381 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4441 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4501 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4561 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4621 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4681 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4741 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4801 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4861 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4921 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4981 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5041 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5101 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5161 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5221 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5281 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5341 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5401 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5461 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5521 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5581 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5641 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5701 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5761 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5821 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5881 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5941 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6001 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6061 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6121 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6181 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6241 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6301 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6361 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6421 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6481 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6541 gctatgacca tgattacgcc aagcgcgccc gccgggtaac tcacggggta tccatgtcca
     6601 tttctgcggc atccagccag gatacccgtc ctcgctgacg taatatccca gcgccgcacc
     6661 gctgtcatta atctgcacac cggcacggca gttccggctg tcgccggtat tgttcgggtt
     6721 gctgatgcgc ttcgggctga ccatccggaa ctgtgtccgg aaaagccgcg acgaactggt
     6781 atcccaggtg gcctgaacga acagttcacc gttaaaggcg tgcatggcca caccttcccg
     6841 aatcatcatg gtaaacgtgc gttttcgctc aacgtcaatg cagcagcagt catcctcggc
     6901 aaactctttc catgccgctt caacctcgcg ggaaaaggca cgggcttctt cctccccgat
     6961 gcccagatag cgccagcttg ggcgatgact gagccggaaa aaagacccga cgatatgatc
     7021 ctgatgcagc taga
//
