LOCUS       sequence-178088-        7297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoterless, Sleeping Beauty-compatible Gateway destination vector,
            IRES Puromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7297)
  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 7297)
  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.IP
FEATURES             Location/Qualifiers
     source          1..7297
                     /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"
     misc_feature    2604..3156
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2748..2765)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2975..2994
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3168..3767
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3168..3187)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3664..3684
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3848..3865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3854..4078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4125..4141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4161..4180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4205..4252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    4393..4514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4430..4449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4563..4579)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4576..4598)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4587..4603
                     /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(4611..4617,4618..4635,4636..4641))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4656..4677
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4794..4811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5271..5859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5351..5370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6030..6821,6822..6890))
                     /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     6653..6672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6891..6995)
                     /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 ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     2641 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     2701 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     2761 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     2821 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     2881 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     2941 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     3001 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     3061 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     3121 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg accgagtaca
     3181 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3241 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3301 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3361 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3421 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3481 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     3541 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     3601 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     3661 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     3721 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     3781 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgatgatc tagagctcgc
     3841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4021 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4081 tctgaggcgg aaagaacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     4141 gtggctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4201 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcatacacgt
     4261 ggtcaggtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4321 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4381 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4501 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4561 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4621 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4681 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4741 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4801 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4861 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg ctcttcgttg
     4921 tggtttgtcc aaactcatca atgtatctta tcatgaatac taattgagtg tatgtaaact
     4981 tctgacccac tgggaatgtg atgaaagaaa taaaagctga aatgaatcat tctctctact
     5041 attattctga tatttcacat tcttaaaata aagtggtgat cctaactgac ctaagacagg
     5101 gaatttttac taggattaaa tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc
     5161 taaggtgtat gtaaacttcc gacttcaact gtatagagta ctgctagcca catgtgagca
     5221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7021 gacgtcgacg tgcgcatcga actccctata cagttgaagt cggaagttta catacactta
     7081 agttggagtc attaaaactc gtttttcaac tactccacaa atttcttgtt aacaaacaat
     7141 agttttggca agtcagttag gacatctact ttgtgcatga cacaagtcat ttttccaaca
     7201 attgtttaca gacagattat ttcacttata attcactgta tcacaattcc agtgggtcag
     7261 aagtttacat acactaagtt gactgtgtta cggcgtc
//
