LOCUS       Exported                7622 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Synaptic Tag for Mapping Activity in GABAergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    mDlx-SynTagMA_post-2A-mCerulean
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  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 7622)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7622
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          849..981
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     964..983
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1293..1316
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1338..1778
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             2595..2936
                     /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             2985..3110
                     /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"
     CDS             3129..3182
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3204..3923
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRNHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3249..3270)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3510..3529)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3857..3878
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3968..4556
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4021..4041)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4439..4450)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4725..4806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4762..4781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4816..4835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4885..5025
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4885..5014
                     /label=AAV2 ITR
     rep_origin      5100..5555
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5187..5206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5397..5418
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5572..5591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5691..5713
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5751..5769)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5837..5941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5942..6802
                     /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(6160..6179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6973..7561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7462..7481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagaattcta tacactcaca gtggtttggc
      181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt
      241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc
      301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc
      361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
      421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
      481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
      541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
      601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
      661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
      781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
      841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaacta
     1021 gcatgctcga agtcaaggaa gcatcaccaa caagcatcca gatcagctgg gtgctccact
     1081 tgcgccacgt tcgctactac cgcatcacct acggtgaaac tggtggcaat agccctgtcc
     1141 aggaattcac cgtgcctggc agcaagtcca ctgctaccat cagcggcctg aaacctggtg
     1201 tcgactatac catcacggtg tacgccgtca cgatcttcag cgcctaccgc tccgcctggc
     1261 cgccgatctc catcaactac cgcaccggaa ccgattacaa ggatgacgac gataagggta
     1321 gcggctccag tagatctgac caactgactg aagagcagat cgcagaattt aaagaggctt
     1381 tctccctatt tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga
     1441 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     1501 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa
     1561 tgaaagacac agacagtgaa gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg
     1621 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     1681 taacagatga agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg
     1741 taaactacga agagtttgta gtgatgatga cagcgaagct cgagtgcgag aaaatctatg
     1801 tgcgtgatgg agtgctgacg ggtgatattc atatgatctt gttgcttgaa ggaaatgccc
     1861 attaccgatg tgacttcaga actacttaca aagctaagga gaagggtgtc aagttaccag
     1921 gcgtgcacta tgtggaccac accattgaga ttttaagcca tgacaaagat tacaacaagg
     1981 ttaagatcta tgagtatgct gttgctcatt ctggattgcc tgacaatgcc agacgaggcg
     2041 gtaccggcgg atccatggtg agtgcgatta agccagacat gaagatcaaa ctgcgtatgg
     2101 aaggcaacgt aaacgggcac cactttgtga tcgacggaga tggtacaggc aagccttatg
     2161 agggaaaaca gaccatggat cttgaagtca aagagggcgg acctctgcct tttgcctttg
     2221 atatcctgac cactgcattc cattacggca acagggtatt cgtgaaatat ccagacaaca
     2281 tacaagacta ttttaagcag tcgtttccta aggggtattc gtgggaacga agcatgactt
     2341 tcgaagacgg gggcatttgc tatgccagaa gcgacataac aatggaaggg gacactttct
     2401 ataataaagt tcgattttat ggtaccaact ttcccgccaa tggtccagtt atgcagaaga
     2461 agacgctgaa atggatgccg agctggacgc gttcatcacg tcgtaagtgg aataagacag
     2521 gtcacgcagt cagagctata ggtcggctgg agctcggagc tggcgctgga gcgggtgcag
     2581 gggctggctc tagattccag tgccggatct gcatgcggaa cttcagcgac cggtccaacc
     2641 tgagcaggca catcagaacc cacaccggag aaaagccctt cgcctgcgac atttgcggcc
     2701 ggaagttcgc catcagcagc aacctgaaca gccacaccaa gatccacact ggcagccaga
     2761 aacctttcca gtgcagaatt tgtatgagaa actttagcag aagcgacaac ctggccagac
     2821 acatccggac acatactggt gaaaaacctt ttgcctgtga tatctgtggc agaaagtttg
     2881 ccacctccgg caatctgacc cggcacacaa agattcacct gcggggcagc cagctatcga
     2941 ttgtcgacgc tcctgaacaa cgtgaaggtg cttctcaagt ttctgtttct gttacttttg
     3001 aagatgttgc tgttcttttt actcgtgatg aatggaaaaa acttgatctt tctcaacgtt
     3061 ctctttatcg tgaagttatg cttgaaaatt attctaatct tgcttctatg gcttacgtac
     3121 ccgcggccga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     3181 ctgcaccggg atccgatatc cccatggtga gcaagggcga ggagctgttc accggggtgg
     3241 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     3301 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     3361 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttcg
     3421 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     3481 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     3541 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3601 aggacggcaa catcctgggg cacaagctgg agtacaacgc catcagcgac aacgtctata
     3661 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     3721 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3781 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     3841 acgagaagcg caatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3901 gcatggacga gctgtacaag taaaggcgcg ccacccctgc agggaattcg atatcaagct
     3961 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4021 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4081 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4141 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4201 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4261 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4321 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4381 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4441 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4501 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4561 ataccgagcg ctgctcgaga gatctgtgat agcggccatc aagctggccg cgactctaga
     4621 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4681 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4741 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4801 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcagcttat cgataccgca
     4861 tgcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4921 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4981 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     5041 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     5101 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5161 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5221 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5281 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5341 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5401 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5461 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5521 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5581 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5641 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5701 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5761 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5821 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5941 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6001 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6061 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6121 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6181 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6241 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6301 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6361 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6421 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6481 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6541 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6601 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6661 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6721 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6781 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6841 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6901 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6961 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7021 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7081 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7141 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7201 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7261 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7321 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7381 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7441 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7501 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7561 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7621 gt
//
