LOCUS       sequence-370666-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector containing the zebrafish ubiquitin B 
            polyadenylation sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Walton EM, Cronan MR, Beerman RW, Tobin DM
  TITLE     The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term 
            Analysis of Macrophage Behavior during Mycobacterial Infection in 
            Zebrafish.
  JOURNAL   PLoS One. 2015 Oct 7;10(10):e0138949. doi: 
            10.1371/journal.pone.0138949. eCollection 2015.
  PUBMED    26445458
REFERENCE   2  (bases 1 to 6157)
  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: "PLoS One.";
            date: "2015-10-7"; volume: "10(10):e0138949. doi"; pages: " 
            10.1371/journal.pone.0138949. eCollection 2015"
COMMENT     Sequence Label: pDEST tol2 UbB polyA
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1220
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1261..1566)
                     /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(1363..1382)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1886..2566)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     2481..2500
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2620..2626,2627..2644,2645..2650))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2675..2798)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3054..3074)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3082..3098)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3114..3248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3138..3157)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3192..3211
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(3687..3874,3875..3886)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3920..3936)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3920..3936)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3973..3993)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3973..3991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4012..4028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4025..4047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4036..4052
                     /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(4060..4066,4067..4084,4085..4090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4105..4126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4243..4260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4414..5002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4494..4513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5173..5964,5965..6033))
                     /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     5796..5815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6034..6138)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttgaac gagaaacgta aaatgatata
     1141 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     1201 caacatatcc agtcactatg gtcgacctgc agactggctg tgtataaggg agcctgacat
     1261 ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc ggtggctgag
     1321 atcagccact tcttccccga taacggagac cggcacactg gccatatcgg tggtcatcat
     1381 gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacggggga ctttatctga
     1441 cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt caataatatc
     1501 actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt aaaccttaaa
     1561 ctgcatttca ccagcccctg ttctcgtcgg caaaggagcc gttcatttca ataaaccggg
     1621 cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc agacgacggg
     1681 cttcattctg catggttgtg cttaccgaac cggagatatt gacatcatat atgccttgag
     1741 caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag catacctctt
     1801 tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa atcagcgcgc
     1861 aaatacgcat actgttatct ggcttttagt aagccggatc ctctagatta cgccccgcct
     1921 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     1981 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     2041 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     2101 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     2161 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     2221 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     2281 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2341 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2401 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2461 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2521 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2581 ctcctgaaaa tctcgacgga tcctaactca aaatccacac attatacgag ccggaagcat
     2641 aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt gactggatat gttgtgtttt
     2701 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2761 attttacgtt tctcgttcaa ctttattata catagttgat attctcagta tcccctgctt
     2821 atgcactcac cactttttcc cattatttta ttcatgttgt gttgctaaat tcaactacat
     2881 tcgtattacc atgttgtttc atttgttttt tttttatctc agctgtcaaa ataaaagcat
     2941 tgaaatacaa cattgttcta tttttcattg tggttttgca tgcccggtaa ataaaaaaac
     3001 aaggtaataa aaataagatg cgaaatgaga agtataattc actggccgtc gttcaacttt
     3061 attatacata gttgataatt cactggccgt cgttttacgg taccatcgat gatgatccag
     3121 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3181 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3241 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3301 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctctag
     3361 atcagatctg cgaagatacg gccacgggtg ctcttgatcc tgtggctgat tttggactgt
     3421 gctgctcgca gctgctgatg aatcacatac ttcctccatt ttcttccact gattgactgt
     3481 tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga
     3541 cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt catataatgt gaaaacgatt
     3601 ttcataggca gaataaataa catttaaatt aaactgggca tcagcgcaat tcaattggtt
     3661 tggtaatagc aagggaaaat agaatgaagt gatctccaaa aaataagtac tttttgactg
     3721 taaataaaat tgtaaggagt aaaaagtact tttttttcta aaaaaatgta attaagtaaa
     3781 agtaaaagta ttgattttta attgtactca agtaaagtaa aaatccccaa aaataatact
     3841 taagtacagt aatcaagtaa aattactcaa gtactttaca cctctggttc ttgaccccct
     3901 accttcagca agcccagcag atccactagt tctagagcgg ccgccaccgc ggtggagctc
     3961 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     4021 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4081 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4141 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4201 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6121 acaaataggg gttccgcgca catttccccg aaaagtg
//
