LOCUS       Exported                6433 bp DNA     circular SYN 20-AUG-2024
DEFINITION  Gateway(R) destination vector for inducible high-level expression of
            proteins in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6433)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6433)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6433)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    67..191
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        338..440
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             441..1097
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1442..1744
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1788..1912)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1990..2037
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2403..2431)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        2544..2648
                     /label=AmpR promoter
     CDS             2649..3506
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3680..4268
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4454..4596)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4701..4889)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagatcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
      121 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      181 cagtcactat ggcggccgct aagttggcag catcacccga cgcactttgc gccgaataaa
      241 tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg
      301 ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca
      361 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
      421 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
      481 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
      541 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
      601 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
      661 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
      721 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      781 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      841 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
      901 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
      961 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1021 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1081 agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg
     1141 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     1201 tgtcaaaaag aggtgtgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     1261 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     1321 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     1381 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga
     1441 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     1501 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     1561 tctgctgtca gataaagtcc cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     1621 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     1681 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     1741 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     1801 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     1861 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtttgattcg
     1921 acccgggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1981 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2041 aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc
     2101 gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag
     2161 tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc
     2221 atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg
     2281 cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc
     2341 attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     2401 gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc
     2461 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2641 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2701 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2761 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2821 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2881 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2941 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3001 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3061 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3121 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3181 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3241 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3301 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3361 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3421 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3481 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3541 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4321 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4381 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4441 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4501 gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4561 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4621 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4681 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4741 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4801 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4861 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     4921 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     4981 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     5041 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     5101 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     5161 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     5221 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     5281 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     5341 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     5401 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     5461 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     5521 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     5581 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     5641 cataaatcgc cgtgacgatc agcggtccag tgatcgaagt taggctggta agagccgcga
     5701 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     5761 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     5821 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     5881 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     5941 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     6001 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6061 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6121 ctgggttgaa ggctctcaag ggcatcggtc gatcgacgct ctcccttatg cgactcctgc
     6181 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     6241 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     6301 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     6361 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     6421 gcgtagagga tcg
//
