LOCUS       sequence-353476-        7129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing piggyBac inverted terminal repeats (ITRs) 
            sequences flanking CAG and Venus .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7129)
  AUTHORS   Yamagata M, Sanes JR
  TITLE     CRISPR-mediated Labeling of Cells in Chick Embryos Based on 
            Selectively Expressed Genes.
  JOURNAL   Bio Protoc. 2021 Aug 5;11(15):e4105. doi: 10.21769/BioProtoc.4105. 
            eCollection 2021 Aug 5.
  PUBMED    34458399
REFERENCE   2  (bases 1 to 7129)
  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: "Bio 
            Protoc."; date: "2021-08-5"; volume: "11(15):e4105. doi"; pages: " 
            10.21769/BioProtoc.4105. eCollection 2021 Aug 5"
COMMENT     Sequence Label: pXL-BacII-CAG-Venus
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..584
                     /label=chicken beta-actin promoter
     intron          585..1601
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1525..1547
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1609..1628
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1668..1692
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1768..1770,1771..1773,1774..2487)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1813..1834)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2074..2093)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2421..2442
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2527..2551)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2642..2661)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2707..2762
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2761..2780)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3385..3419
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3910..3926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3910..3926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3923..3945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3934..3950
                     /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)."
     promoter        complement(join(3958..3964,3965..3982,3983..3988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4003..4024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4141..4158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4312..4900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4392..4411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5071..5862,5863..5931))
                     /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     5694..5713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5932..6036)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6062..6517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6199..6220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6411..6430
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6644..6666
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6658..6675
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6659..6675
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6805..6867
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      421 gggggggggc gcgcccgggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      481 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      541 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      601 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      661 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      721 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      781 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      841 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      901 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      961 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     1021 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     1081 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     1141 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     1201 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     1261 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     1321 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     1381 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1441 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1501 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1561 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1621 ttattgtgct gtctcatcat tttggcaaag aattcctcga ggatatcaca agtttgtaca
     1681 aaaaagcagg ctccgaattc gccctttctc gagctcaagc ttcgaattct gcagtcgacg
     1741 gtaccgcggg cccgggatcg atccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1801 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1861 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1921 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc
     1981 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2041 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2101 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2161 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2221 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2281 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2341 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     2401 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2461 ctcggcatgg acgagctgta caagtaagaa ttggccgcga ctctagatca taaaagggcg
     2521 aattcgaccc agctttcttg tacaaagtgg tgattcgcgg ccgcactcct caggtgcagg
     2581 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     2641 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     2701 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2761 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     2821 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     2881 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     2941 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     3001 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     3061 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt gatatctata
     3121 acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata acaatataat
     3181 tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca
     3241 cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg
     3301 agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa
     3361 agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt
     3421 taatctagct gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa
     3481 gctggcgcta tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag
     3541 cggcatggaa agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca
     3601 cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg
     3661 ttcaggccac ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca
     3721 gcccgaagcg catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg
     3781 tgcagattaa tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg
     3841 gctggatgcc gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc
     3901 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3961 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4021 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4501 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4741 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4861 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4921 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4981 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5041 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5161 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5221 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5281 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5581 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5941 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6001 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6061 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     6121 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     6181 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     6241 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     6301 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     6361 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     6421 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     6481 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     6541 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     6601 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     6661 aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg
     6721 cagactcgcg gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg
     6781 ctgcagaaca cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa
     6841 gataatcatg tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta
     6901 ttaatttgaa tagatattaa gttttattat atttacactt acatactaat aataaattca
     6961 acaaacaatt tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc
     7021 tataaagtaa caaaactttt atcgaattcc tgcagcccgg gggatccact agttattaat
     7081 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
