LOCUS       sequence-241696-        6963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Global labeling of inhibitory synapses (red fluorescence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  AUTHORS   Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff 
            SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man HY, Han X
  TITLE     A Viral Toolbox of Genetically Encoded Fluorescent Synaptic Tags.
  JOURNAL   iScience. 2020 Jun 30;23(7):101330. doi: 10.1016/j.isci.2020.101330.
  PUBMED    32674057
REFERENCE   2  (bases 1 to 6963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience.";
            date: "2020-06-30"; volume: "23(7):101330. doi"; pages: " 
            10.1016/j.isci.2020.101330"
COMMENT     Sequence Label: AAV-EF1A-mScarlet-Gephyrin.FingR-IL2RGTC
FEATURES             Location/Qualifiers
     source          1..6963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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)."
     primer_bind     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   380..509
                     /label=AAV2 ITR
     promoter        598..1776
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          829..1767
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1724..1744
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1793..1812
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1793..1811
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1838..1847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1844..2539
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1977..1995)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2245..2264
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2846..2869
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3287..3412
                     /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    3472..4060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3525..3545)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3943..3954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4063..4079)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4064..4080)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4109..4230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4120..4139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4174..4193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4392..4521
                     /label=AAV2 ITR
     primer_bind     complement(4540..4557)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4540..4556)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4549..4571)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4698..5153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4785..4804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4995..5016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5179..5283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5284..5352,5353..6144)
                     /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(5502..5521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6315..6903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6804..6823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatcac gcgtcttcca cagagtgaat tccgtgaggc
      601 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      661 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      721 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      781 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      841 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      901 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
      961 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     1021 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     1081 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     1141 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     1201 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     1261 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     1321 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     1381 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     1441 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1501 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1561 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     1621 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     1681 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     1741 gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa ttagcttggt actaatacga
     1801 ctcactatag ggagacccaa gctggctagg taagcttgcc accatggtga gcaagggcga
     1861 ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca tgaacggcca
     1921 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1981 gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt cccctcagtt
     2041 catgtacggc tccagggcct tcatcaagca ccccgccgac atccccgact actataagca
     2101 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgccgt
     2161 gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg tgaagctccg
     2221 cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg gctgggaagc
     2281 gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta agatggccct
     2341 gcgcctgaag gacggcggcc gctacctggc ggacttcaag accacctaca aggccaagaa
     2401 gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca cctcccacaa
     2461 cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact ccaccggcgg
     2521 catggacgag ctgtacaagt ctaggggagc tggcgctgga gcgggtgcag gggctggcct
     2581 cgaagtcaag gaagcatcac caaccagcat ccagatcagc tggggcaagt acaaggtcat
     2641 ggttcgctac taccgcatca cctacggtga aactggtggc aatagccctg tccaggaatt
     2701 caccgtgcct ggcagcaagt ccactgctac catcagcagc ctgaaacctg gtgtcgacta
     2761 taccatcacg gtgtacgccg tcacgatcga ccactggaac taccaggacc cgatcccgat
     2821 ctccatcaac taccgcaccg gatccgatta caaggatgac gacgataagg gtagcggctc
     2881 cagtagatct gggtttcagt gccagatttg catgcgcaac tttagccgca aaagcaccct
     2941 gaccgatcat attcgcaccc ataccggcga aaaaccgttt gcgtgcgata tttgcggccg
     3001 caaatttgcg gcgcgcagca cccgcaccac ccataccaaa attcataccg gcagccagaa
     3061 accgtttcag tgccgcattt gcatgcgcaa ctttagccgc agcgatagcc tgagcaaaca
     3121 tattcgcacc cataccggcg aaaaaccgtt tgcgtgcgat atttgcggcc gcaaatttgc
     3181 gcagcgcagc aacctgaaag tgcataccaa aattcatctg cgcggcagcc agctgatcga
     3241 tggtgtcgac gctcctgaac aacgtgaagg tgcttctcaa gtttctgttt ctgttacttt
     3301 tgaagatgtt gctgttcttt ttactcgtga tgaatggaaa aaacttgatc tttctcaacg
     3361 ttctctttat cgtgaagtta tgcttgaaaa ttattctaat cttgcttcta tggcttaaaa
     3421 gcttgcggcc gcgactctag agtcgacctg caggcatgca agcttatcga taatcaacct
     3481 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3541 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3601 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3661 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3721 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3781 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3841 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     3901 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3961 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4021 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     4081 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4141 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4201 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4261 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4321 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     4381 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4441 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4501 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     4561 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4621 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4681 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4741 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4801 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4861 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4921 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4981 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5041 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5101 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5161 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5521 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5821 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6421 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6961 atg
//
