LOCUS       sequence-178090-        7751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoterless, Sleeping Beauty-compatible Gateway destination vector,
            Zeocin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7751)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 7751)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pSBDEST.Z
FEATURES             Location/Qualifiers
     source          1..7751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     210..229
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        210..228
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    258..382
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(811..1116)
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(913..932)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1436..2119)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2034..2053
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2120..2222)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2400..2524)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2550..2591
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2631..2650)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2675..2723
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    2907..3028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             3091..3132
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3142..3159
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3182..3199)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3188..3412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3458..3886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3545..3564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3755..3776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3895..3915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3900..4229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4080..4215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4142..4161
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4277..4324
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4343..4717
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQMVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4847..4968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5017..5033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5017..5033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5030..5052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5041..5057
                     /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(5065..5071,5072..5089,5090..5095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5110..5131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5248..5265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5805..5824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6484..7275,7276..7344))
                     /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     7107..7126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7345..7449)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattctctg gctaactaga
      181 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
      241 ggctagttaa gctatcaaca agtttgtaca aaaaagctga acgagaaacg taaaatgata
      301 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
      361 cacaacatat ccagtcacta tgaatcaact acttagatgg tattagtgac ctgtagtcga
      421 ccgacagcct tccaaatgtt cttcgggtga tgctgccaac ttagtcgacc gacagccttc
      481 caaatgttct tctcaaacgg aatcgtcgta tccagcctac tcgctattgt cctcaatgcc
      541 gtattaaatc ataaaaagaa ataagaaaaa gaggtgcgag cctctttttt gtgtgacaaa
      601 ataaaaacat ctacctattc atatacgcta gtgtcatagt cctgaaaatc atctgcatca
      661 agaacaattt cacaactctt atacttttct cttacaagtc gttcggcttc atctggattt
      721 tcagcctcta tacttactaa acgtgataaa gtttctgtaa tttctactgt atcgacctgc
      781 agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc
      841 gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac
      901 cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac
      961 cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac
     1021 catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg
     1081 gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag
     1141 caaaagagcc gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg
     1201 ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccagac
     1261 cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac
     1321 tgtaatacgc tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta
     1381 tatattctta taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt
     1441 aagccggatc cacgcggcgt ttacgccccc cctgccactc atcgcagtac tgttgtaatt
     1501 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1561 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     1621 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     1681 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     1741 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     1801 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     1861 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     1921 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     1981 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2041 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2101 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2161 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     2221 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     2281 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt
     2341 cgggtgatgc tgccaactta gtcgactaca ggtcactaat accatctaag tagttgattc
     2401 atagtgactg gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
     2461 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag
     2521 tggttgatct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     2581 tcgattctac gcgtaccggt tagtaatgag tttaaacggg ggaggctaac tgaaacacgg
     2641 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     2701 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     2761 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     2821 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     2881 tgccatagca gatcttatta aagcagaact tgtttattgc agcttataat ggttacaaat
     2941 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3001 gtttgtccaa actcatcaat gtatcttatc atgtctggtc gaatccatca cactggcggc
     3061 cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt
     3121 ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     3181 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3241 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3301 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3361 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3421 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3481 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3541 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3601 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3661 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3721 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3781 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3841 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3901 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3961 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4081 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4141 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4201 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4261 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4321 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     4381 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     4441 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     4501 ccaggaccag atggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     4561 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     4621 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     4681 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     4741 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4801 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4861 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4921 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4981 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5041 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5101 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5161 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5341 ggataacgca ggaaagaaca tgtgctcttc gttgtggttt gtccaaactc atcaatgtat
     5401 cttatcatga atactaattg agtgtatgta aacttctgac ccactgggaa tgtgatgaaa
     5461 gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc acattcttaa
     5521 aataaagtgg tgatcctaac tgacctaaga cagggaattt ttactaggat taaatgtcag
     5581 gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac ttccgacttc
     5641 aactgtatag agtactgcta gccacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacgtgcgca tcgaactccc
     7501 tatacagttg aagtcggaag tttacataca cttaagttgg agtcattaaa actcgttttt
     7561 caactactcc acaaatttct tgttaacaaa caatagtttt ggcaagtcag ttaggacatc
     7621 tactttgtgc atgacacaag tcatttttcc aacaattgtt tacagacaga ttatttcact
     7681 tataattcac tgtatcacaa ttccagtggg tcagaagttt acatacacta agttgactgt
     7741 gttacggcgt c
//
