LOCUS       sequence-334783-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway destination vector to create transgenes for 
            phiC31-mediated transgenesis in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  AUTHORS   Mosimann C, Puller AC, Lawson KL, Tschopp P, Amsterdam A, Zon LI
  TITLE     Site-directed zebrafish transgenesis into single landing sites with 
            the phiC31 integrase system.
  JOURNAL   Dev Dyn. 2013 Aug;242(8):949-63. doi: 10.1002/dvdy.23989. Epub 2013 
            Jul 3.
  PUBMED    23723152
REFERENCE   2  (bases 1 to 5083)
  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: 
            "10.1002/dvdy.23989"; journalName: "Dev Dyn"; date: "2013-08"; 
            volume: "242"; issue: "8"; pages: "949-63"
COMMENT     Sequence Label: pDestattB (pCM268)
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     primer_bind     801..817
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    837..960
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1001..1306)
                     /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(1103..1122)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1626..2306)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     2221..2240
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2360..2366,2367..2384,2385..2390))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2415..2538)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2546..2562)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2607..2676
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(2895..2913)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2934..2950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2947..2969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2958..2974
                     /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(2982..2988,2989..3006,3007..3012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3027..3048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3165..3182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3336..3924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3416..3435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4095..4886,4887..4955))
                     /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     4718..4737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4956..5060)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatctag cggatccggg aattgggaat tcgttaacag
      781 atctgcggcc gcggctcgag caggaaacag ctatgaccat gattacgcca agctatcaac
      841 tttgtataga aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta
      901 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg
      961 gtcgacctgc agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca
     1021 ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga
     1081 taacggagac cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga
     1141 tatgcaccac cgggtaaagt tcacggggga ctttatctga cagcagacgt gcactggcca
     1201 gggggatcac catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca
     1261 gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagcccctg
     1321 ttctcgtcgg caaaggagcc gttcatttca ataaaccggg cgacctcagc catcccttcc
     1381 tgattttccg ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg
     1441 cttaccgaac cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt
     1501 caactgtcac tgtaatacgc tgcttcatag catacctctt tttgacatac ttcgggtata
     1561 catatcagta tatattctta taccgcaaaa atcagcgcgc aaatacgcat actgttatct
     1621 ggcttttagt aagccggatc ctctagatta cgccccgcct gccactcatc gcagtactgt
     1681 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1741 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1801 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1861 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1921 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1981 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     2041 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga
     2101 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     2161 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     2221 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     2281 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgacgga
     2341 tcctaactca aaatccacac attatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2401 cctaatgcgg ccgccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2461 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa
     2521 ctttattata catagttgat aattcactgg ccgtcgtttt acggtacctc tagagatcca
     2581 ctagtgtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc
     2641 ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat gaacgggtcg
     2701 aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa
     2761 ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat
     2821 acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgac actagttcta
     2881 gccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     2941 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3001 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3061 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3121 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3181 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3241 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3301 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3781 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3961 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4021 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4081 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4141 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4201 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4261 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4321 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4381 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4441 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4501 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4561 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4621 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4681 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4741 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4801 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4861 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4921 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4981 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5041 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
