LOCUS       Exported                6680 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Synaptic Tag for Mapping Activity.
ACCESSION   .
VERSION     .
KEYWORDS    SynTagMA_post
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6680)
  AUTHORS   Perez-Alvarez A, Fearey BC, O'Toole RJ, Yang W, Arganda-Carreras I, 
            Lamothe-Molina PJ, Moeyaert B, Mohr MA, Panzera LC, Schulze C, 
            Schreiter ER, Wiegert JS, Gee CE, Hoppa MB, Oertner TG
  TITLE     Freeze-frame imaging of synaptic activity using SynTagMA.
  JOURNAL   Nat Commun. 2020 May 18;11(1):2464. doi: 10.1038/s41467-020-16315-4.
  PUBMED    32424147
REFERENCE   2  (bases 1 to 6680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6680
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        23..470
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             760..783
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             805..1245
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             2062..2403
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     CDS             2452..2577
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     misc_feature    2596..3184
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2649..2669)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3067..3078)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3187..3203)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3188..3204)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3271..3747
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3470..3489)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3787..3927
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3787..3916
                     /label=AAV2 ITR
     rep_origin      4002..4457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4089..4108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4299..4320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4474..4493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4593..4615
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4653..4671)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4739..4843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4844..5704
                     /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     complement(5062..5081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5875..6463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6364..6383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6525..6654
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gctgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg
       61 ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc
      121 aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg
      181 cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc
      241 gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc
      301 ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc
      361 gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc
      421 ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgaattcaa
      481 gctgctagca tgctcgaagt caaggaagca tcaccaacaa gcatccagat cagctgggtg
      541 ctccacttgc gccacgttcg ctactaccgc atcacctacg gtgaaactgg tggcaatagc
      601 cctgtccagg aattcaccgt gcctggcagc aagtccactg ctaccatcag cggcctgaaa
      661 cctggtgtcg actataccat cacggtgtac gccgtcacga tcttcagcgc ctaccgctcc
      721 gcctggccgc cgatctccat caactaccgc accggaaccg attacaagga tgacgacgat
      781 aagggtagcg gctccagtag atctgaccaa ctgactgaag agcagatcgc agaatttaaa
      841 gaggctttct ccctatttga caaggacggg gatgggacaa taacaaccaa ggagctgggg
      901 acggtgatgc ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat
      961 gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgac aatgatggca
     1021 agaaaaatga aagacacaga cagtgaagaa gaaattagag aagcgttccg tgtgtttgat
     1081 aaggatggca atggctacat cagtgcagca gagcttcgcc acgtgatgac aaaccttgga
     1141 gagaagttaa cagatgaaga ggttgatgaa atgatcaggg aagcagacat cgatggggat
     1201 ggtcaggtaa actacgaaga gtttgtagtg atgatgacag cgaagctcga gtgcgagaaa
     1261 atctatgtgc gtgatggagt gctgacgggt gatattcata tgatcttgtt gcttgaagga
     1321 aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa gggtgtcaag
     1381 ttaccaggcg tgcactatgt ggaccacacc attgagattt taagccatga caaagattac
     1441 aacaaggtta agatctatga gtatgctgtt gctcattctg gattgcctga caatgccaga
     1501 cgaggcggta ccggcggatc catggtgagt gcgattaagc cagacatgaa gatcaaactg
     1561 cgtatggaag gcaacgtaaa cgggcaccac tttgtgatcg acggagatgg tacaggcaag
     1621 ccttatgagg gaaaacagac catggatctt gaagtcaaag agggcggacc tctgcctttt
     1681 gcctttgata tcctgaccac tgcattccat tacggcaaca gggtattcgt gaaatatcca
     1741 gacaacatac aagactattt taagcagtcg tttcctaagg ggtattcgtg ggaacgaagc
     1801 atgactttcg aagacggggg catttgctat gccagaagcg acataacaat ggaaggggac
     1861 actttctata ataaagttcg attttatggt accaactttc ccgccaatgg tccagttatg
     1921 cagaagaaga cgctgaaatg gatgccgagc tggacgcgtt catcacgtcg taagtggaat
     1981 aagacaggtc acgcagtcag agctataggt cggctggagc tcggagctgg cgctggagcg
     2041 ggtgcagggg ctggctctag attccagtgc cggatctgca tgcggaactt cagcgaccgg
     2101 tccaacctga gcaggcacat cagaacccac accggagaaa agcccttcgc ctgcgacatt
     2161 tgcggccgga agttcgccat cagcagcaac ctgaacagcc acaccaagat ccacactggc
     2221 agccagaaac ctttccagtg cagaatttgt atgagaaact ttagcagaag cgacaacctg
     2281 gccagacaca tccggacaca tactggtgaa aaaccttttg cctgtgatat ctgtggcaga
     2341 aagtttgcca cctccggcaa tctgacccgg cacacaaaga ttcacctgcg gggcagccag
     2401 ctatcgattg tcgacgctcc tgaacaacgt gaaggtgctt ctcaagtttc tgtttctgtt
     2461 acttttgaag atgttgctgt tctttttact cgtgatgaat ggaaaaaact tgatctttct
     2521 caacgttctc tttatcgtga agttatgctt gaaaattatt ctaatcttgc ttctatggct
     2581 taaaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2641 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2701 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2761 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2821 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2881 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2941 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3001 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3061 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3121 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3181 ccgcatcgat accgtcgacc tcgagacgcg atcgcggcgc gccattcgat ggcctctctg
     3241 gcctcgacct cgagtctaga gatgatctac gggtggcatc cctgtgaccc ctccccagtg
     3301 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3361 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3421 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3481 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3541 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3601 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3661 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3721 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3781 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3841 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3901 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3961 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4021 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4081 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4141 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4201 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4261 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4321 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4381 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4441 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4501 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4561 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4621 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4681 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4741 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4801 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4861 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4921 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4981 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5041 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5101 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5161 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5221 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5281 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5341 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5401 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5461 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5521 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5581 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5641 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5701 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5761 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5821 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6001 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     6541 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     6601 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     6661 gcacgcgtcg ctagactcga
//
