LOCUS       Exported                4693 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  VAMP2 tagged with SNAP.
ACCESSION   .
VERSION     .
KEYWORDS    VAMP2-SNAP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4693)
  AUTHORS   Martineau M, Somasundaram A, Grimm JB, Gruber TD, Choquet D, Taraska
            JW, Lavis LD, Perrais D
  TITLE     Semisynthetic fluorescent pH sensors for imaging exocytosis and 
            endocytosis.
  JOURNAL   Nat Commun. 2017 Nov 10;8(1):1412. doi: 10.1038/s41467-017-01752-5.
  PUBMED    29123102
REFERENCE   2  (bases 1 to 4693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4693
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             1178..1723
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             1768..1791
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1806..1940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1830..1849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1884..1903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2051..2067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2051..2067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2064..2086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2075..2091
                     /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)."
     promoter        complement(2099..2129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2144..2165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2282..2299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2453..3041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2533..2552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3212..4072)
                     /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     3835..3854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4073..4177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4245..4263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4301..4323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4423..4442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4636..4658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4650..4667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4651..4667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggccca ggcccgaatt catgtcggct accgctgcca ccgtcccgcc
      841 tgccgccccg gccggcgagg gtggcccccc tgcacctcct ccaaatctta ccagtaacag
      901 gagactgcag cagacccagg cccaggtgga tgaggtggtg gacatcatga gggtgaatgt
      961 ggacaaggtc ctggagcgag accagaagct atcggaactg gatgatcgcg cagatgccct
     1021 ccaggcaggg gcctcccagt ttgaaacaag tgcagccaag ctcaagcgca aatactggtg
     1081 gaaaaacctc aagatgatga tcatcttggg agtgatttgc gccatcatcc tcatcatcat
     1141 catcgtttac ttcagcactg gcggatccgg cggatccatg gacaaagatt gcgaaatgaa
     1201 acgtaccacc ctggatagcc cgctgggcaa actggaactg agcggctgcg aacagggcct
     1261 gcatgaaatt aaactgctgg gtaaaggcac cagcgcggcc gatgcggttg aagttccggc
     1321 cccggccgcc gtgctgggtg gtccggaacc gctgatgcag gcgaccgcgt ggctgaacgc
     1381 gtattttcat cagccggaag cgattgaaga atttccggtt ccggcgctgc atcatccggt
     1441 gtttcagcag gagagcttta cccgtcaggt gctgtggaaa ctgctgaaag tggttaaatt
     1501 tggcgaagtg attagctatc agcagctggc ggccctggcg ggtaatccgg cggccaccgc
     1561 cgccgttaaa accgcgctga gcggtaaccc ggtgccgatt ctgattccgt gccatcgtgt
     1621 ggttagctct agcggtgcgg ttggcggtta tgaaggtggt ctggcggtga aagagtggct
     1681 gctggcccat gaaggtcatc gtctgggtaa accgggtctg ggacctgcag gtataggcgc
     1741 gccaggatcc taactcgagg taccggcgac tacaaggacg acgatgacaa gtaagcggcc
     1801 gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1861 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1921 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1981 gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg
     2041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3181 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3241 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3301 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3361 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3421 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3481 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3541 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3601 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3661 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3721 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3781 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3841 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3901 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3961 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4021 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4081 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4141 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4201 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4261 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4321 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4381 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4441 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4501 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4561 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4621 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
     4681 catgcctgca ggt
//
