LOCUS       Exported                6918 bp ds-DNA     circular SYN 01-MAR-2021
DEFINITION  SNAP25 FRET Kinetics substrate.
ACCESSION   urn.local...1g-3js4sas
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 2, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6918
                     /organism="synthetic DNA construct"
     source          137..157
     source          158..178
     source          158..178
     source          158..178
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          179..904
     source          179..874
     source          179..874
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          875..904
     source          875..904
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          905..922
     source          923..1120
     source          923..1120
     source          join(923..1120,923..1120)
     source          join(923..1120,923..1120)
     source          1121..1135
     source          1136..1849
     source          1136..1849
     source          join(1136..1849,1136..1849)
     misc_feature    69..87
                     /label=t7 term
                     /note="t7 term"
                     /note="Geneious type: terminator"
     CDS             complement(137..1849)
                     /standard_name="CV-SNAP25"
                     /label=CV-SNAP25
     misc_feature    complement(137..157)
                     /standard_name="Hisx6"
                     /label=Hisx6
     misc_feature    complement(158..874)
                     /standard_name="mVenus"
                     /label=mVenus
     primer_bind     complement(200..221)
                     /standard_name="EGFP-C"
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     549..568
                     /standard_name="EXFP-R"
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     808..829
                     /standard_name="EGFP-N"
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(881..904)
                     /codon_start=1
                     /standard_name="Xpress(TM) tag"
                     /product="Xpress(TM) epitope tag, including anenterokinase 
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /note="Geneious type: misc_feature"
                     /translation="DLYDDDDK"
     misc_feature    complement(923..1120)
                     /standard_name="SNAP25(141-206)"
                     /label=SNAP25(141-206)
     misc_feature    complement(1136..1849)
                     /standard_name="eCFP"
                     /label=eCFP
     promoter        complement(1918..1935)
                     /standard_name="T7 promoter"
                     /label=T7 promoter
     CDS             2325..3404
                     /standard_name="LacI"
                     /label=LacI
     unsure          2879..2884
     unsure          3120..3125
     rep_origin      complement(4819..5438)
                     /direction=LEFT
     misc_feature    5544..6359
                     /standard_name="KAN"
                     /label=KAN
                     /note="Geneious type: ORF"
     CDS             5544..6359
                     /standard_name="KanR"
                     /label=KanR
     rep_origin      complement(6453..6908)
                     /direction=LEFT
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctt gtacagctcg tccatgccga
      181 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
      241 ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc
      301 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga
      361 tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat
      421 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
      481 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
      541 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
      601 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
      661 agcactgcag gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
      721 cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
      781 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
      841 ccccggtgaa cagctcctcg cccttgctca ccatcggatc cttatcgtca tcgtcgtaca
      901 gatcacttcc accactattt gacccagacc ccagcatttt ggtcgcacgt tggttagcct
      961 cgtcaatgcg tgtcttattg ctatcagcct tttccatgat gcgatcaatc tgacgatttt
     1021 gggtatcaat ttcattgccc atatctaatg ccatatgacg caagttgcca atgatcccag
     1081 atacctgctc aaggttctca tccatttcat tttcgcgggc acgactacgc ccacctttat
     1141 acagttcatc cataccatga gtaataccag cggcggtaac gaactctaaa aggaccatat
     1201 ggtcgcgttt ctcattgggg tccttagaca gggctgactg tgtgctcaaa tagtgattat
     1261 caggtaaaag gacggggcca tctccaattg gtgtgttttg ctggtaatgg tctgcaagct
     1321 ggacgcttcc atcctcaatg ttgtgacgaa tcttgaaatt tgccttgatt ccgttttttt
     1381 gtttatcagc cgtgatgtag acgttatgag agatatagtt gtattctaat ttatgtccta
     1441 agatgttgcc atcctcttta aagtcgatac ccttcaactc aatgcgatta acaagagtat
     1501 ctccctcaaa cttaacctcg gcgcgggtct tatagtttcc atcatcttta aaaaaaatgg
     1561 tgcgttcctg gacgtaacct tctggcatcg cggacttgaa aaagtcgtgt tgtttcatat
     1621 ggtcagggta gcgagcgaaa cactgcacgc cccaggcgaa cgtagtaacc aatgtaggcc
     1681 acggtactgg aagcttgccg gtagtgcaaa taaattttaa tgtcaacttt ccgtacgtcg
     1741 catcgccctc tccttcccca gatacggaaa acttgtgacc attcacatcg ccatccagtt
     1801 ctacaagaat ggggaccacg ccggtaaaca attcctcccc cttagacatg tatatctcct
     1861 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
     1921 tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg gacgcatcgt
     1981 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     2041 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     2101 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     2161 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     2221 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
     2281 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     2341 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     2401 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     2461 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     2521 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     2581 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     2641 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     2701 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     2761 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     2821 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     2881 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     2941 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     3001 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     3061 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     3121 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     3181 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     3241 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     3301 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3361 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3421 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     3481 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     3541 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     3601 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     3661 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     3721 gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     3781 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     3841 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     3901 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     3961 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     4021 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     4081 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     4141 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     4201 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     4261 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     4321 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc
     4381 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4441 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4501 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     4561 cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag
     4621 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     4681 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4741 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg tctgcttaca
     5521 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctctaggc
     5581 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     5641 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     5701 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     5761 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     5821 atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat
     5881 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     5941 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     6001 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     6061 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     6121 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     6181 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     6241 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     6301 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaag
     6361 aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6421 ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa
     6481 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6541 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6601 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6661 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6721 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6781 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6841 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6901 ggcgcgtccc attcgcca
//
