LOCUS       sequence-316182-        8363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-iCre-ERT2 piggyBac transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     piggyBac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8363)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PB CAG-iCre-ERT2
FEATURES             Location/Qualifiers
     source          1..8363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1739..1748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2792..3727
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2840..3733
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     misc_feature    3770..4358
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3823..3843)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4241..4252)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4397..4414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4403..4627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4981..5015
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5102..5121)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5103..5121)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5128..5145)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5128..5144)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5137..5159)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5286..5741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5423..5444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5635..5654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5768..5872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5873..5941,5942..6733)
                     /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(6091..6110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6904..7492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7393..7412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7646..7663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7780..7801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7816..7821,7822..7839,7840..7846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7854..7870
                     /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     7859..7881
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7878..7894
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7878..7894
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7913..7933
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7915..7933
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   7997..8059
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggsgcgcgc cggggggggg gggggggggg gggggggggg
      541 ggggggsgrg gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg ctagcgccgc
     1741 caccatgggc gccagcaacc tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga
     1801 tgccacctct gatgaagtca ggaagaacct gatggacatg ttcagggaca ggcaggcctt
     1861 ctctgaacac acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa
     1921 gctgaacaac aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta
     1981 cctgcaagcc agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat
     2041 gctgcacagg agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat
     2101 gaggagaatc agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt
     2161 tgaacgcact gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga
     2221 catcaggaac ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat
     2281 tgccagaatc agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat
     2341 tggcaggacc aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt
     2401 taccaagctg gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta
     2461 cctgttctgc cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc
     2521 cacccgggcc ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga
     2581 tgactctggg cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccag
     2641 ggacatggcc agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa
     2701 tgtgaacatt gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag
     2761 gctgctcgag gatggggacc tcgagccatc tgctggagac atgagagctg ccaacctttg
     2821 gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt ccctgacggc
     2881 cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt ccgagtatga
     2941 tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc tggcagacag
     3001 ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg atttgaccct
     3061 ccatgatcag gtccaccttc tagaatgtgc ctggctagag atcctgatga ttggtctcgt
     3121 ctggcgctcc atggagcacc cagtgaagct actgtttgct cctaacttgc tcttggacag
     3181 gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc tggctacatc
     3241 atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca aatctattat
     3301 tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc tggaagagaa
     3361 ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc tgatggccaa
     3421 ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc tcatcctctc
     3481 ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga agtgcaagaa
     3541 cgtggtgccc ctctatgacc tgctgctgga ggcggcggac gcccaccgcc tacatgcgcc
     3601 cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg ccactgcggg
     3661 ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag agggtttccc
     3721 tgccacagct tgattaatta atgatcatac agtgctctga gtacgctata atcaacctct
     3781 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3841 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3901 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3961 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4021 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4081 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4141 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     4201 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4261 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4321 tcagacgagt cggatctccc tttgggccgc ctccccgcag gatccactag agcggccgcc
     4381 tagagctcgc tatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4441 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4501 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4561 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4621 tctatggctt ctgaggcgga aagaaccagc tggggctcga ctagagcttg cggaaccctt
     4681 aagctttttt attatatatt agtcacgata tctataacaa gaaaatatat atataataag
     4741 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
     4801 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
     4861 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
     4921 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
     4981 catgcgtcaa ttttacgcag actatctttc tagggttaaa aaagatttgc gctttactcg
     5041 acctaaactt taaacacgtc atagaatctt cgtttgacaa aaaccacatt gtgttgggtc
     5101 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     5161 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5221 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5281 atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5341 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5401 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5461 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5521 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5581 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5641 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5701 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     5761 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5821 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5881 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5941 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6001 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6061 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6121 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6181 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6241 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6301 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6361 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6421 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6481 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6541 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6601 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6661 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6721 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6781 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6841 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6901 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6961 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7021 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7081 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7141 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7201 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7261 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7321 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7381 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7441 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7501 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7561 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7621 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7681 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7741 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7801 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7861 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     7921 ccctcactaa agggaacctc ccccggtatt cacgacagca ggctgaataa taaaaaaatt
     7981 agaaactatt atttaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
     8041 tgcgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
     8101 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
     8161 tttatttatg tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag
     8221 taacaaaact tttaaacatt ctctctttta caaaaataaa cttattttgt actttaaaaa
     8281 cagtcatgtt gtattataaa ataagtaatt agcttaacct atacataata gaaacaaatt
     8341 atacttatta accggtcatg gtc
//
