LOCUS       Exported                6900 bp ds-DNA     circular SYN 01-MAR-2021
DEFINITION  VAMP4 FRET Kinetics substrate.
ACCESSION   urn.local...1g-3js4sas
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  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..6900
                     /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..1102
     source          923..1102
     source          1103..1117
     source          1118..1831
     source          1118..1831
     source          join(1118..1831,1118..1831)
     misc_feature    69..87
                     /label=t7 term
                     /note="t7 term"
                     /note="Geneious type: terminator"
     CDS             complement(137..1831)
                     /standard_name="CV-VAMP4"
                     /label=CV-VAMP4
     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..1102)
                     /standard_name="VAMP4 (P48–R107)"
                     /label=VAMP4 (P48–R107)
     misc_feature    complement(1118..1831)
                     /standard_name="eCFP"
                     /label=eCFP
     promoter        complement(1900..1917)
                     /standard_name="T7 promoter"
                     /label=T7 promoter
     CDS             2307..3386
                     /standard_name="LacI"
                     /label=LacI
     unsure          2861..2866
     unsure          3102..3107
     rep_origin      complement(4801..5420)
                     /direction=LEFT
     misc_feature    5526..6341
                     /standard_name="KAN"
                     /label=KAN
                     /note="Geneious type: ORF"
     CDS             5526..6341
                     /standard_name="KanR"
                     /label=KanR
     rep_origin      complement(6435..6890)
                     /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 gagcggcgca gttgcttact acgattagag aaggcggtgg
      961 cattgtctga aagagattca ctcttgtcct gtaattcatc aaggcgctct ccacgttcaa
     1021 ttaccttagt aatgttctcc tgcattacgt caatcacttc gtctacttga ttttggacgt
     1081 gtttgatttt gtcgttacgt ggacgactac gcccaccttt atacagttca tccataccat
     1141 gagtaatacc agcggcggta acgaactcta aaaggaccat atggtcgcgt ttctcattgg
     1201 ggtccttaga cagggctgac tgtgtgctca aatagtgatt atcaggtaaa aggacggggc
     1261 catctccaat tggtgtgttt tgctggtaat ggtctgcaag ctggacgctt ccatcctcaa
     1321 tgttgtgacg aatcttgaaa tttgccttga ttccgttttt ttgtttatca gccgtgatgt
     1381 agacgttatg agagatatag ttgtattcta atttatgtcc taagatgttg ccatcctctt
     1441 taaagtcgat acccttcaac tcaatgcgat taacaagagt atctccctca aacttaacct
     1501 cggcgcgggt cttatagttt ccatcatctt taaaaaaaat ggtgcgttcc tggacgtaac
     1561 cttctggcat cgcggacttg aaaaagtcgt gttgtttcat atggtcaggg tagcgagcga
     1621 aacactgcac gccccaggcg aacgtagtaa ccaatgtagg ccacggtact ggaagcttgc
     1681 cggtagtgca aataaatttt aatgtcaact ttccgtacgt cgcatcgccc tctccttccc
     1741 cagatacgga aaacttgtga ccattcacat cgccatccag ttctacaaga atggggacca
     1801 cgccggtaaa caattcctcc cccttagaca tgtatatctc cttcttaaag ttaaacaaaa
     1861 ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
     1921 tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     1981 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     2041 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     2101 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     2161 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     2221 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     2281 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     2341 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     2401 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     2461 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     2521 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     2581 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     2641 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     2701 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     2761 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     2821 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     2881 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     2941 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     3001 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     3061 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     3121 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     3181 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     3241 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     3301 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3361 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     3421 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3481 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3541 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     3601 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     3661 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     3721 gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3781 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3841 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3901 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     3961 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     4021 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     4081 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     4141 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     4201 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     4261 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     4321 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4381 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4441 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4501 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4561 tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4621 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4681 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4741 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4801 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4861 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4921 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4981 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5041 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5101 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5161 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5221 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5281 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5341 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5401 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5461 ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     5521 gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca
     5581 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     5641 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     5701 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     5761 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     5821 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     5881 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     5941 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     6001 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     6061 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     6121 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     6181 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     6241 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     6301 ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga
     6361 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6421 aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     6481 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6541 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6601 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6661 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6721 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6781 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6841 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
