LOCUS       sequence-333794-        6441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cargo vector for the phiC31 system with, unc-119 rescue fragment, no
            fluorescent marker. Gateway R3R4 sites (for gene of interest 
            insertion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  AUTHORS   Yang FJ, Chen CN, Chang T, Cheng TW, Chang NC, Kao CY, Lee CC, Huang
            YC, Hsu JC, Li J, Lu MJ, Chan SP, Wang J
  TITLE     phiC31 integrase for recombination-mediated single-copy insertion 
            and genome manipulation in Caenorhabditis elegans.
  JOURNAL   Genetics. 2022 Feb 4;220(2):iyab206. doi: 10.1093/genetics/iyab206.
  PUBMED    34791215
REFERENCE   2  (bases 1 to 6441)
  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: "Genetics.";
            date: "2022-02-4"; volume: "220(2):iyab206. doi"; pages: " 
            10.1093/genetics/iyab206"
COMMENT     Sequence Label: pCG150_double_phiC31_attB
FEATURES             Location/Qualifiers
     source          1..6441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..184
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     169..185
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2396..2429
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     2575..2597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2635..2653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2721..2825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2826..2894,2895..3686)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3044..3063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3857..4445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4346..4365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4507..4540
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     4621..4643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4640..4656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4640..4656
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4676..4799
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4840..5145)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(4942..4961)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5465..6145)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     6060..6079
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6199..6205,6206..6223,6224..6229))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6254..6377)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6385..6402)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6385..6401)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6394..6416)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
       61 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      121 ccttacgcat ctgtgcggta tttcacaccg catatgccgg gcccccctcg aggtcgacgg
      181 tatcgataag cttcagtaaa agaagtagaa ttttatagtt ttttttctgt ttgaaaaatt
      241 ctccccatca atgttctttc aaataaatac atcactaatg caaagtattc tataacctca
      301 tatctaaatt cttcaaaatc ttaacatatc ttatcattgc tttaagtcaa acgtaacatt
      361 aaaaaaaatg ttttggaaaa tgtgtcaagt ctctcaaaat tcagtttttt aaaccactcc
      421 tatagtccta tagtcctata gttacccatg aaatccttat atattactgt aaaatgtttc
      481 aaaaaccatt ggcaaattgc cagaactgaa aatttccggc aaattgggga accggcaaat
      541 tgccaatttg ctgaatttgc cggaaaaacg gtaattgccg aaagtttttg acacgaaaat
      601 ggcaaattgt ggttttaaaa tttttttttt tggaaatttc agaatttcaa ttttaatcgg
      661 caaaactgta ggcatcctaa gaatgttcct acatctattt tgaaaagtaa gcgaattcta
      721 tgaaaatgtc taaagaaaat ggggaaacaa tttcaaaaag gcacagtttc aagtgtttcc
      781 gaattatact aaatccctct aaaaacttcc ggcaaattga tatccgtaaa agagcaaatc
      841 cgcatttttg ccgaaaatta aaatttccga caaatcggca aaccggcaat ttggcgaaat
      901 ttgccggaag gcaattgccg cccacccctg ttccagaggt tcaaactggt agcaaagctc
      961 aaaatttctc aaattctcca attttttttt gaattttggc agtgtaccaa aatgacattc
     1021 agtcatattg gtttattata gatttattta gataaaatcc taaatgattc tacctttaaa
     1081 gatgcccact ttaaaagtaa tgactcaaac ttcaaattgc tctaagattc tattgaatta
     1141 ccatcttttc ctctcatttt ctctcactgt ctatttcatc acaaattcat ccctctctcc
     1201 ctctctttct ctctccctct ctctctttct ctttgctcat catctgtcat tttgtccgtt
     1261 cctctctctg cgccctcagc gttccccaca ctctctcgct tctcttttcc tagacgtctt
     1321 cttttttcat cttcttcagc cctttttcgc cattttccat ctctgtcaat cattacggac
     1381 gacccccatt aatttttgga tgtgctctgc gagcaagagc acacggaact catttatttt
     1441 ttgttggaat ttctttttca ttttagttag ttaaaagcta caacagcata tatgaaggca
     1501 gagcaacaac aacaatcgat cgcacccggc tcggcaacct tcccgtctca gatgccccga
     1561 cccccaccgg taaccgaaca ggctataacc accgaggcgg agcttctcgc gaaaaatcaa
     1621 attacaccaa atgatgtgct agctcttcca ggaatcactc aaggattcct atgctcccca
     1681 tcggcaaacg tctataacat tgagttcacc aaattccaaa tccgtgacct cgacacggaa
     1741 cacgtgctct tcgaaattgc aaaaccggaa aatgagacgg aagagaatct gcaggcgcaa
     1801 gccgaatcgg caagatatgt ccgatatcga tttgcgccga attttctgaa attaaagacg
     1861 gtcggcgcga cggtcgagtt caaggtcggc gatgtgccga tcacacattt tcgaatgatt
     1921 gaacggcact ttttcaagga tcgtttattg aaatgctttg attttgaatt tggattctgt
     1981 atgccaaatt cgagaaacaa ctgtgaacat atctatgaat ttccacaact ttcacaacaa
     2041 ctcatggacg acatgatcaa taatccaaac gagacccgct ccgatagctt ctattttgtg
     2101 gagaataagc tcgtaatgca caataaggcc gactactcgt atgatgcata atttcccgcc
     2161 gatggaataa aactaataat ttttctcgca atttatagaa atcatgtgat tttcctaccg
     2221 ctaaatttaa aatttcctaa tttttcacta tatttctcac aaaatgcatt ccgaaaactc
     2281 tcatttttca gcttttcaat acaaatccca ataaatgatt tttcaaatat aaaaaaaaaa
     2341 acggaattcc tgcagcccgg gggatccgct agttctagag cggccgccac cgctacgcgc
     2401 ccggggagcc caagggcacg ccctggcacc cagctcccat atggtgcact ctcagtgcaa
     2461 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2521 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2581 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2641 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2701 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2761 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2821 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2881 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2941 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3001 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3061 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3121 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3181 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3241 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3301 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3361 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3421 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3481 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3541 ggtctcgcgg tatcattgca gcactggggt cagatggtaa gccctcccgt atcgtagtta
     3601 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3661 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3721 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3781 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3841 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3901 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3961 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4021 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4081 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4141 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4201 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4261 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4321 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4381 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4441 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg ccttttgctc
     4501 acatgggtgc cagggcgtgc ccttgggctc cccgggcgcg tacctgcagg catgttcttt
     4561 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4621 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctatcaact
     4681 ttgtatagaa aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     4741 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     4801 tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc agaacatcag
     4861 gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat
     4921 aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat
     4981 atgcaccacc gggtaaagtt cacgggggac tttatctgac agcagacgtg cactggccag
     5041 ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag
     5101 acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt
     5161 tctcgtcggc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct
     5221 gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc
     5281 ttaccgaacc ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc
     5341 aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact tcgggtatac
     5401 atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg
     5461 gcttttagta agccggatcc tctagattac gccccgcctg ccactcatcg cagtactgtt
     5521 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     5581 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     5641 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     5701 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     5761 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     5821 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     5881 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
     5941 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     6001 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     6061 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     6121 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacggat
     6181 cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc
     6241 ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     6301 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     6361 tttattatac atagttgata attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     6421 ccctggcgtt acccaactta a
//
