LOCUS       sequence-181065-        8262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector for bacterial expression; His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8262)
  TITLE     Gateway Destination vectors for E. coli expression
REFERENCE   2  (bases 1 to 8262)
  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"
COMMENT     Sequence Label: pDest-527
FEATURES             Location/Qualifiers
     source          1..8262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2..20
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    21..45
                     /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)."
     RBS             60..82
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             105..122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    135..259
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(284..289,290..307,308..314)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             368..1027
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(434..453)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1080..1668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2300..2605
                     /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     2484..2503
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2646..2770)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2830..2862
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2869..2886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3052..3138
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3230..3257
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3311..3329)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3325..3372
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3409..3864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3496..3515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3706..3727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3979..4770,4771..4839))
                     /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     4602..4621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4840..4932)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5013..5601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5787..5926
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5852..5871)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6012..6034
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6028..6219)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6791..7873)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7835..7854
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7874..7951)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8157..8176
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
       61 ttgtttaact ttaagaagga gatatacata tgagatctgg atcccaccat caccatcacc
      121 atagatctga tatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
      181 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
      241 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
      301 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
      361 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      421 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
      481 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
      541 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
      601 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
      661 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
      721 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
      781 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
      841 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
      901 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
      961 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1021 ggcgtaaacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     1861 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1921 cccgactgga aagcgggcag tgagcgaacg caattaatac gcgtggatcc ggcttactaa
     1981 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     2041 atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac
     2101 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     2161 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     2221 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     2281 gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     2341 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     2401 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
     2461 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
     2521 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     2581 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg
     2641 tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     2701 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt
     2761 acaaagtggt gatatcgcta gctagatagc tagaagcttg cggccgcaca gctgtataca
     2821 cgtgcaagcc agccagaact cgctcctgaa gacccagagg atctcgagca ccaccaccac
     2881 caccactaat gttaattaag ttgggcgttc ctaggctgat aaaacagaat ttgcctggcg
     2941 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     3001 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     3061 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3121 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     3181 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     3241 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     3301 aaatatgtat ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     3361 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     3421 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3481 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3541 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3601 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3661 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3721 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3781 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3841 acaaaatatt aacgtttaca atttctggcg gcacgatggc atgagattat caaaaaggat
     3901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg
     4861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4921 aaaataaaca aataggtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4981 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5041 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5101 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     5161 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5221 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5281 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5341 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5401 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5461 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5521 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5581 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5641 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5701 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5761 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5821 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5881 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     5941 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6001 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6061 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     6121 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     6181 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     6241 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     6301 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     6361 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     6421 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     6481 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     6541 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     6601 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     6661 caggagcacg atcatgctag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     6721 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     6781 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     6901 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     6961 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     7021 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     7081 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     7141 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     7201 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     7261 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     7321 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     7381 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     7441 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     7501 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     7561 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     7621 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     7681 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     7741 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     7801 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     7861 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     7921 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     7981 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     8041 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     8101 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     8161 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     8221 cgtccggcgt agaggatcga gatcgatctc gatcccgcga aa
//
