pAAV-Syn-hCRAFi-CCR2_P2A_Fusionred

LOCUS       pAAV-Syn-hCRAFi-CCR2_P2A_Fusionred               7285 bp DNA     circular SYN 07-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7285)
  AUTHORS   Kiryl Piatkevich
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    311..800
                     /label=Syn
     promoter        349..796
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     709..728
                     /label=21-HSYN-F
     regulatory      809..820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             821..2644
                     /codon_start=1
                     /product="Isoform B"
                     /label=hCCR2B
                     /translation="MLSTSRSRFIRNTNESGEEVTTFFDYDYGAPCHKFDVKQIGAQLL
                     PPLYSLVFIFGFVGNMLVVLILINCKKLKCLTDIYLLNLAISDLLFLITLPLWAHSAAN
                     EWVFGNAMCKLFTGLYHIGYFGGIFFIILLTIDRYLAIVHAVFALKARTVTFGVVTSVI
                     TWLVAVFASVPGIIFTKCQKEDSVYVCGPYFPRGWNNFHTIMRNILGLVLPLLIMVICY
                     SGILKTLLRCRNGLEEGGNVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIG
                     DGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSK
                     GEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVT
                     TLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVN
                     RIELKGIDFKEDGNILGHKLEYNIDKKRHRAVRVIFTIMIVYFLFWTPYNIVILLNTFQ
                     EFFGLSNCESTSQLDQATQVTETLGMTHCCINPIIYAFVGEKFRRYLSVFFRKHITKRF
                     CKQCPVFYRETVDGVTSTNTPSTGEQEVSAGL"
     CDS             965..967
                     /codon_start=1
                     /label=Y49A mutation
                     /translation="Y"
     CDS             1178..1180
                     /codon_start=1
                     /label=Y120A
                     /translation="Y"
     primer_bind     1343..1370
                     /label=415-1173-F1
     CDS             1541..2263
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=cpGFP
                     /translation="NVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYN"
     primer_bind     1885..1902
                     /label=309-WKG1111-F
     CDS             2438..2440
                     /codon_start=1
                     /label=T292A
                     /translation="T"
     primer_bind     2629..2648
                     /label=416-1175-F
     CDS             2660..2719
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             2720..2779
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             2780..2839
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             2846..2866
                     /codon_start=1
                     /label=ER2
                     /translation="FCYENEV"
     CDS             2891..2947
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2960..3655
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     3280..3300
                     /label=111-WKG-368-F3
     misc_feature    3683..4271
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4154..4165)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4289..4496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4548..4688
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4763..5218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5500..5604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5605..6465
                     /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"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aattaattaa tggttaatta atctagactg cagagggccc tgcgtatgag tgcaagtggg
      361 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      421 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      481 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      541 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      601 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      661 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      721 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      781 gtgcctgaga gcgcagtcga gaggatccgg taccgccacc atgctgagca ccagcagaag
      841 cagattcatc agaaacacca acgagagcgg cgaggaggtg accaccttct tcgactacga
      901 ctacggcgcc ccctgccaca agttcgacgt gaagcagatc ggcgcccagc tgctgccccc
      961 cctgtacagc ctggtgttca tcttcggctt cgtgggcaac atgctggtgg tgctgatcct
     1021 gatcaactgc aagaagctga agtgcctgac cgacatctac ctgctgaacc tggccatcag
     1081 cgacctgctg ttcctgatca ccctgcccct gtgggcccac agcgccgcca acgagtgggt
     1141 gttcggcaac gccatgtgca agctgttcac cggcctgtac cacatcggct acttcggcgg
     1201 catcttcttc atcatcctgc tgaccatcga cagatacctg gccatcgtgc acgccgtgtt
     1261 cgccctgaag gccagaaccg tgaccttcgg cgtggtgacc agcgtgatca cctggctggt
     1321 ggccgtgttc gccagcgtgc ccggcatcat cttcaccaag tgccagaagg aggacagcgt
     1381 gtacgtgtgc ggcccctact tccccagagg ctggaacaac ttccacacca tcatgagaaa
     1441 catcctgggc ctggtgctgc ccctgctgat catggtgatc tgctacagcg gcatcctgaa
     1501 gaccctgctg agatgcagaa acggcctgga ggagggagga aacgtctata tcaaggccga
     1561 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcgg
     1621 cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1681 gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg
     1741 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1801 gctgtacaag ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg
     1861 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1921 cggcgagggt gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1981 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2041 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2101 aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2161 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2221 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacatcgaca agaagagaca
     2281 cagagccgtg agagtgatct tcaccatcat gatcgtgtac ttcctgttct ggacccccta
     2341 caacatcgtg atcctgctga acaccttcca ggagttcttc ggcctgagca actgcgagag
     2401 caccagccag ctggaccagg ccacccaggt gaccgagacc ctgggcatga cccactgctg
     2461 catcaacccc atcatctacg ccttcgtggg cgagaagttc agaagatacc tgagcgtgtt
     2521 cttcagaaag cacatcacca agagattctg caagcagtgc cccgtgttct acagagagac
     2581 cgtggacggc gtgaccagca ccaacacccc cagcaccggc gagcaggagg tgagcgccgg
     2641 cctgatggac gagctgtaca agagtagaat cacaagcgaa ggcgagtaca tccccctgga
     2701 tcaaatagac ataaatgtaa agagtagaat cacaagcgaa ggcgagtaca tccccctgga
     2761 tcaaatagac ataaatgtaa agagtagaat cacaagcgaa ggcgagtaca tccccctgga
     2821 tcaaatagac ataaatgtag gtggattttg ttatgagaat gaagtgcgac cggttcacgt
     2881 gggaagtggg gctacaaact tctcactgct gaaacaggcc ggggatgtgg aggaaaaccc
     2941 aggaccacac gtgactagta tggtgagcga gctgattaag gagaacatgc ccatgaagct
     3001 gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa
     3061 gccctacgag ggcacccaga ccatgagaat caaggtcgtc gagggcggcc ctctcccctt
     3121 cgccttcgac atcctggcta ccagcttcat gtacggcagc agaaccttca tcaagcaccc
     3181 tccgggcatc cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt
     3241 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
     3301 ctgcctcatc tacaacgtca aggttagagg ggtgaacttc ccagccaacg gccctgtgat
     3361 gcagaagaaa acactcggct gggaggcctc caccgagacg atgtaccctg ctgacggagg
     3421 actggaaggc gcatgtgaca tggctctgaa gctcgtggga ggaggacacc tgatctgcaa
     3481 ccttgagaca acatacagat ccaagaaacc cgctacgaac ctcaagatgc ccggcgtcta
     3541 caacgtggac cacagactgg aaagaatcaa ggaggccgac gatgagacat acgtcgagca
     3601 gcacgaggtg gctgtggcca gatactctac tggtggcgct ggtgatggag gtaaataaga
     3661 attccgtaca aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3721 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3781 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3841 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3901 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3961 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4021 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4081 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4141 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4201 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4261 cgcctccccg catcgatacc ggctcgagct gtgccttcta gttgccagcc atctgttgtt
     4321 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4381 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4441 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4501 gtctcgagct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4561 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4621 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4681 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4741 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4801 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4861 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4921 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4981 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5041 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     5101 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     5161 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     5221 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5281 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5341 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5401 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5461 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5521 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5581 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5641 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5701 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5761 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5821 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5881 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5941 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6001 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6061 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6121 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6181 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6241 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6301 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6361 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6421 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7261 ttttgctggc cttttgctca catgt
//