LOCUS       sequence-82396-e        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for PhiC31 integrase-mediated cassette exchange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Tasic B, Hippenmeyer S, Wang C, Gamboa M, Zong H, Chen-Tsai Y, Luo L
  TITLE     Site-specific integrase-mediated transgenesis in mice via pronuclear
            injection.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 10;108(19):7902-7. doi: 
            10.1073/pnas.1019507108. Epub 2011 Apr 4.
  PUBMED    21464299
REFERENCE   2  (bases 1 to 6249)
  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"; doi: 
            "10.1073/pnas.1019507108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-05-10- 10"; volume: "108"; issue: "19"; pages: "7902-7"
COMMENT     Sequence Label: pBT378_pattB-pCA-GFP-pA-attB
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(624..1415,1416..1484))
                     /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     1247..1266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1485..1589)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1615..2070)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1752..1773)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1964..1983
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2197..2219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2211..2228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2212..2228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2235..2254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2235..2253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2304..2373
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     enhancer        2679..2982
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2984..3261
                     /label=chicken beta-actin promoter
     intron          3263..4280
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4204..4226
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4288..4307
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4358..4367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4364..4366,4367..4369,4370..5083)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4409..4430)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4670..4689)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5017..5038
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5205..5326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5242..5261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5296..5315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    5389..5458
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(5673..5691)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5712..5728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5712..5728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5725..5747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5736..5752
                     /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(5760..5766,5767..5784,5785..5790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5805..5826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5943..5960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6114..6249,1..453))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6194..6213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
       61 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      301 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     1621 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1681 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1741 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1801 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1861 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1921 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1981 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2041 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     2101 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     2161 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     2221 cggccagtga attgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg
     2281 aggtcgacga tgtaggtcac ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg
     2341 ggctccccgg gcgcgtactc cacctcaccc atctggtcca tcatgatgaa cgggtcgagg
     2401 tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg
     2461 ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg
     2521 cggggcagcg tcagcgggtt ctcgacggtc acggcgggca tgtcgacggt atcgataagc
     2581 ttcccttaat taaccgttta aacaattctg caggaatcta gttattaata gtaatcaatt
     2641 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2701 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2761 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2821 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2881 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2941 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     3001 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     3061 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
     3121 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
     3181 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
     3241 aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc gtgccccgct
     3301 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     3361 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     3421 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     3481 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     3541 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     3601 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     3661 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     3721 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     3781 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     3841 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     3901 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     3961 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     4021 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     4081 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     4141 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     4201 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     4261 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     4321 ttggcaaaga attgatttga taccgcgggc ccgggccgcc accatggtga gcaagggcga
     4381 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4441 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4501 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4561 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4621 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4681 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4741 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4801 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt
     4861 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     4921 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4981 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     5041 cgccgccggg atcactctcg gcatggacga gctttacaag taagaattct agataactga
     5101 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5161 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttattta aattgggcgc
     5341 gcccaactag ttctagagcg gccgccactc gacgatgtag gtcacggtct cgaagccgcg
     5401 gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccacct cacccatctg
     5461 gtccatcatg atgaacgggt cgaggtggcg gtagttgatc ccggcgaacg cgcggcgcac
     5521 cgggaagccc tcgccctcga aaccgctggg cgcggtggtc acggtgagca cgggacgtgc
     5581 gacggcgtcg gcgggtgcgg atacgcgggg cagcgtcagc gggttctcga cggtcacggc
     5641 gggcatgtcg agtggagctc cagcttttgt tccctttagt gagggttaat ttcgagcttg
     5701 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5761 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5821 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5881 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5941 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6001 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6061 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6121 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6181 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6241 gttccgacc
//
