LOCUS       Exported                6117 bp ds-DNA     circular SYN 11-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6117
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        25..43
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             75..97
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             105..758
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    792..816
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             837..857
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(843..866)
                     /label=pDEST15_Vector_rev
     misc_feature    867..1554
                     /label=Spt6 1223-1451
                     /note="note=""direct strand 
                     translation""translation=""YYFPFNGRQAEDYLRSKERGEFVIRQSSRGDD
                     HLVITWKLDKDLFQHIDIQELEKENPLALGKVLIVDNQKYNDLDQIIVEYLQNKVRLLN
                     EMTSSEKFKSGTKKDVVKFIEDYSRVNPNKSVYYFSLNHDNPGWFYLMFKINANSKLYT
                     WNVKLTNTGYFLVNYNYPSVIQLCNGFKTLLKSNSSKNRMNNYR*"""
     primer_bind     complement(1018..1050)
                     /label=Spt6_R1282H_QC_2_rev
     primer_bind     complement(1018..1046)
                     /label=Spt6_R1282H_QC_rev
     primer_bind     1040..1067
                     /label=Spt6_R1282H_QC_2_for
     primer_bind     1044..1067
                     /label=Spt6_R1282H_QC_for
     primer_bind     complement(1234..1269)
                     /label=Spt6_K1355A_QC_2_rev
     primer_bind     complement(1234..1265)
                     /label=Spt6_K1355A_QC_rev
     primer_bind     1259..1293
                     /label=Spt6_K1355A_QC_2_for
     primer_bind     1263..1293
                     /label=Spt6_K1355A_QC_for
     primer_bind     complement(1474..1509)
                     /label=Spt6_K1435A_QC_2_rev
     primer_bind     complement(1474..1505)
                     /label=Spt6_K1435A_QC_rev
     primer_bind     1499..1544
                     /label=Spt6_K1435A_QC_2_for
     primer_bind     1503..1544
                     /label=Spt6_K1435A_QC_for
     primer_bind     1554..1572
                     /label=pDEST15_Vector_for
     primer_bind     complement(1569..1579)
                     /label=pDEST15_Vector_for
     protein_bind    complement(1576..1600)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1664..1682
                     /label=T7term
                     /note="note=""direct strand 
                     translation""
translation=""PLSNN*"""
     terminator      1678..1725
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2091..2119)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2232..2336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2337..3197
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3368..3956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4142..4284
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4386..4577)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5787..5796)
                     /label=pDEST15_Vector_for
ORIGIN
        1 atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
       61 cctctagaaa taattttgtt taactttaag aaggagatat acatatgtcc cctatactag
      121 gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag
      181 aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt
      241 ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa
      301 cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc
      361 caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg
      421 tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc
      481 tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg
      541 atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg
      601 acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta
      661 tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct
      721 ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctggtt ccgcgtccat
      781 ggtcgaatca aacaagtttg tacaaaaaag caggctccgc ggccgccgta ctcgaggaaa
      841 acctgtattt tcagggctcc ttcaccagca tttgggactt gaaacaagaa ctggaagatg
      901 ccgaagagga gaggaaattg atgatggcag aagcccgtgc aaagagaaca catcgtgtta
      961 tcaatcatcc ttactatttc cctttcaacg gcagacaggc tgaggattac ttaaggagta
     1021 aagaacgtgg tgaattcgtg atccaccagt ctagccgagg tgatgaccac ttggttatca
     1081 cctggaaatt ggataaggat ttgtttcaac atattgatat ccaagaatta gaaaaagaaa
     1141 atcctttggc tttaggtaaa gtcttgattg tcgacaatca gaaatacaat gatttagacc
     1201 agatcattgt agaatatctt caaaacaagg taaggctctt gaatgaaatg acatctagtg
     1261 aagcattcaa aagcggtact aagaaagatg tggtcaagtt tattgaagac tactctagag
     1321 tgaatccaaa taagtctgtg tactatttca gtttgaacca cgataaccct ggttggtttt
     1381 acttgatgtt caagattaac gcaaatagca aattatacac atggaatgtg aaattaacga
     1441 acactggtta tttcctggta aactacaatt atccaagtgt tatccagctt tgtaatggtt
     1501 ttgcgacgct tctaaaatct aacagtagta agaatagaat gaacaactac cgttagtaga
     1561 agggtgggcg cgccgaccca gctttcttgt acaaagtggt ttgattcgac ccgggatccg
     1621 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1681 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1741 atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc
     1801 aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac
     1861 gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc
     1921 gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
     1981 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
     2041 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
     2101 atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc
     2161 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2341 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2401 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2461 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2521 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     2581 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2641 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2701 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2761 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2821 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2881 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2941 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3001 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3061 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3121 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3481 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4021 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4081 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4141 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
     4201 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4261 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4321 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4381 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     4441 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4501 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4561 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4621 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4681 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4741 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4801 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4861 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4921 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4981 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     5041 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     5101 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     5161 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     5221 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     5281 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     5341 tgacgatcag cggtccagtg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     5401 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     5461 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     5521 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     5581 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     5641 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     5701 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5761 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5821 ctctcaaggg catcggtcga tcgacgctct cccttatgcg actcctgcat taggaagcag
     5881 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5941 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     6001 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     6061 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagagg
//
