LOCUS       sequence-457887-        5995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A CMV-driven Phi31 recombinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5995)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 5995)
  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: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pCS2-phiC31o (JDW 453)
FEATURES             Location/Qualifiers
     source          1..5995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      133..142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             139..1953
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     CDS             1954..1974
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1978..2002)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2047..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2125..2144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2294..2314)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2294..2312)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2333..2349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2333..2349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2346..2368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2357..2373
                     /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(2381..2387,2388..2405,2406..2411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2426..2447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2564..2581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2735..3323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2815..2834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3494..4285,4286..4354))
                     /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     4117..4136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4355..4459)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4485..4940)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4622..4643)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4834..4853
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5011..5995
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctgccaccat ggatacctac gccggagcct acgacagaca gagccgggag
      181 agagagaaca gcagcgccgc cagccccgcc acccagagaa gcgccaacga ggataaggcc
      241 gccgatctgc agagagaggt ggagagggac ggcggcagat tcagatttgt gggccacttc
      301 agcgaggccc ctggcaccag cgccttcggc accgccgaga gacccgagtt cgagagaatc
      361 ctgaacgagt gtagggccgg caggctgaac atgatcatcg tgtacgacgt gtcccggttc
      421 agcaggctga aggtgatgga cgccatccct atcgtgtccg agctgctggc cctgggcgtg
      481 accatcgtgt ccacccagga aggcgtcttt agacagggca acgtgatgga cctgatccac
      541 ctgatcatga ggctggacgc cagccacaag gagagcagcc tgaagagcgc caagatcctg
      601 gacaccaaga acctgcagag ggagctgggc ggctatgtgg gcggcaaggc cccctacggc
      661 ttcgagctgg tgtccgagac caaggagatc acccggaacg gcaggatggt gaacgtggtg
      721 atcaacaagc tggcccacag caccaccccc ctgaccggcc ccttcgagtt tgagcccgac
      781 gtgatcaggt ggtggtggcg ggagatcaag acccacaagc acctgccttt caagcccggc
      841 agccaggccg ccatccaccc cggcagcatc accggcctgt gtaagagaat ggacgccgac
      901 gccgtgccca ccagaggcga gaccatcggc aagaaaaccg ccagcagcgc ctgggacccc
      961 gccaccgtga tgagaatcct gagggaccct aggatcgccg gcttcgccgc cgaggtgatc
     1021 tacaagaaga agcccgacgg cacccccacc accaagatcg agggctacag aatccagaga
     1081 gaccccatca ccctgagacc tgtggagctg gactgtggcc ctatcatcga gcctgccgag
     1141 tggtacgagc tgcaggcctg gctggacggc agaggcagag gcaagggcct gagcagaggc
     1201 caggccatcc tgagcgccat ggacaagctg tactgtgagt gtggcgccgt gatgaccagc
     1261 aagagaggcg aggagagcat caaggacagc taccggtgcc ggagaagaaa ggtggtggac
     1321 cccagcgccc ctggccagca cgagggcacc tgtaatgtga gcatggccgc cctggacaag
     1381 ttcgtggccg agcggatctt caacaagatc cggcacgccg agggcgacga ggagaccctg
     1441 gccctgctgt gggaggccgc cagaagattc ggcaagctga ccgaggcccc cgagaagagc
     1501 ggcgagaggg ccaacctggt ggccgagaga gccgacgccc tgaacgccct ggaggagctg
     1561 tacgaggaca gagccgccgg agcctatgac ggccctgtgg gcaggaagca cttcagaaag
     1621 cagcaggccg ccctgaccct gagacagcag ggcgccgagg aaagactggc cgagctggag
     1681 gccgccgagg cccctaagct gcccctggat cagtggttcc ccgaggatgc cgacgccgac
     1741 cccaccggcc ccaagtcctg gtggggcaga gccagcgtgg acgacaagag ggtgttcgtg
     1801 ggcctgttcg tggataagat cgtggtgacc aagagcacca ccggcagggg ccagggcacc
     1861 cccatcgaga agagagccag catcacctgg gccaagcctc ccaccgacga cgacgaggat
     1921 gacgcccagg acggcaccga ggacgtggcc gcccctaaga aaaagcggaa agtgtgaacc
     1981 cagctttctt gtacaaagtg gttgatcctc tcgagcctct agaactatag tgagtcgtat
     2041 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2101 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2161 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2221 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     2281 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     2341 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     2401 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2461 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2521 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2581 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2641 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3241 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3301 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3361 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3421 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3481 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3541 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3601 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3661 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3721 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3781 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3841 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3901 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3961 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4021 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4081 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4141 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4201 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4261 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4321 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4381 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4441 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4501 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4561 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4621 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4681 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4741 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4801 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4861 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4921 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4981 atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg
     5041 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
     5101 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     5161 gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt
     5221 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     5281 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     5341 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     5401 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     5461 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     5521 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     5581 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     5641 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     5701 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     5761 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca
     5821 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     5881 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     5941 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
