pAAV-GfaABC1D_hCRAFi-CCR2null

LOCUS       pAAV-GfaABC1D_hCRAFi-CCR2null             7153 bp DNA     circular SYN 25-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          995..1127
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1168..1226
                     /label=gfa_NheI_hR2_Fw
     promoter        1172..1190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1207..1212
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1213..3036
                     /codon_start=1
                     /product="Isoform B"
                     /label=hCCR2B
                     /translation="MLSTSRSRFIRNTNESGEEVTTFFDYDYGAPCHKFDVKQIGAQLL
                     PPLASLVFIFGFVGNMLVVLILINCKKLKCLTDIYLLNLAISDLLFLITLPLWAHSAAN
                     EWVFGNAMCKLFTGLAHIGYFGGIFFIILLTIDRYLAIVHAVFALKARTVTFGVVTSVI
                     TWLVAVFASVPGIIFTKCQKEDSVYVCGPYFPRGWNNFHTIMRNILGLVLPLLIMVICY
                     SGILKTLLRCRNGLEEGGNVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIG
                     DGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSK
                     GEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVT
                     TLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVN
                     RIELKGIDFKEDGNILGHKLEYNIDKKRHRAVRVIFTIMIVYFLFWTPYNIVILLNTFQ
                     EFFGLSNCESTSQLDQATQVTETLGMTHCCINPIIYAFVGEKFRRYLSVFFRKHITKRF
                     CKQCPVFYRETVDGVTSTNTPSTGEQEVSAGL"
     primer_bind     1213..1235
                     /label=R2_Fw
     primer_bind     complement(1339..1384)
                     /label=Y49A_Rv_new
     misc_feature    1357..1359
                     /label=Y49A
     primer_bind     1360..1384
                     /label=Y49A_new_Fw
     primer_bind     1471..1491
                     /label=sequencing primer
     primer_bind     complement(1548..1593)
                     /label=Y120A_Rv_new
     misc_feature    1570..1572
                     /label=Y120A
     primer_bind     1573..1593
                     /label=Y120A_new_Fw
     primer_bind     1834..1851
                     /label=NEW SEQUENCING PRIMER
     primer_bind     complement(1896..1938)
                     /label=R2_LEGG_Rv
     primer_bind     complement(1902..1959)
                     /label=R2-3_L1_IO_2aa_Rv
     primer_bind     complement(1904..1959)
                     /label=R2-3_L1_IO_3aa_Rv
     primer_bind     complement(1904..1915)
                     /label=NT_SV_GFP_Erro_RV
     primer_bind     1918..1959
                     /label=LEGG_GFP_Fw
     CDS             1921..1932
                     /codon_start=1
                     /product="Isoform B"
                     /label=hCCR2B
                     /translation="EEGG"
     CDS             1933..2655
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=cpGFP
                     /translation="NVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYN"
     primer_bind     3002..3058
                     /label=HCCR2b  citrine_KGCX3 Fw
     primer_bind     complement(3002..3058)
                     /label=引物 HCCR2b  citrine_KGCX3 Rv
     CDS             3052..3111
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     complement(3084..3111)
                     /label=KGC_GG_Rv
     CDS             3112..3171
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     complement(3144..3171)
                     /label=KGC_GG_Rv
     CDS             3172..3231
                     /codon_start=1
                     /label=KGC
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     complement(3204..3237)
                     /label=KGC_GG_Rv
     primer_bind     complement(3228..3286)
                     /label=gfap_ER2_EcoRV_Rv
     primer_bind     complement(3228..3286)
                     /label=gfabc_KGC_ER2_EcoRV_Rv
     primer_bind     complement(3232..3261)
                     /label=GGER2_NotI_Rv
     CDS             3238..3258
                     /codon_start=1
                     /label=ER2
                     /translation="FCYENEV"
     misc_feature    3280..3868
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3751..3762)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3816..3827
                     /label=Y49A_new_Fw
     polyA_signal    3900..4376
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4416..4556
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4631..5086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5368..5472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5473..6333
                     /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      6504..7092
                     /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 gcgttgtagt taatgattaa cccgccatgc tacttatcta
      181 cgtagccatg ctctaggaag atctaacata tcctggtgtg gagtagggga cgctgctctg
      241 acagaggctc gggggcctga gctggctctg tgagctgggg aggaggcaga cagccaggcc
      301 ttgtctgcaa gcagacctgg cagcattggg ctggccgccc cccagggcct cctcttcatg
      361 cccagtgaat gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt
      421 cccgccgcac cccagccccc ctcaaatgcc ttccgagaag cccattgagc agggggcttg
      481 cattgcaccc cagcctgaca gcctggcatc ttgggataaa agcagcacag ccccctaggg
      541 gctgcccttg ctgtgtggcg ccaccggcgg tggagaacaa ggctctattc agcctgtgcc
      601 caggaaaggg gatcagggga tgcccaggca tggacagtgg gtggcagggg gggagaggag
      661 ggctgtctgc ttcccagaag tccaaggaca caaatgggtg aggggagagc tctccccata
      721 gctgggctgc ggcccaaccc caccccctca ggctatgcca gggggtgttg ccaggggcac
      781 ccgggcatcg ccagtctagc ccactccttc ataaagccct cgcatcccag gagcgagcag
      841 agccagagca ggttggagag gagacgcatc acctccgctg ctcgcaagct ttattgcggt
      901 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
      961 aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1021 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1081 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1141 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga
     1201 gaattcgcca ccatgctgag caccagcaga agcagattca tcagaaacac caacgagagc
     1261 ggcgaggagg tgaccacctt cttcgactac gactacggcg ccccctgcca caagttcgac
     1321 gtgaagcaga tcggcgccca gctgctgccc cccctggcca gcctggtgtt catcttcggc
     1381 ttcgtgggca acatgctggt ggtgctgatc ctgatcaact gcaagaagct gaagtgcctg
     1441 accgacatct acctgctgaa cctggccatc agcgacctgc tgttcctgat caccctgccc
     1501 ctgtgggccc acagcgccgc caacgagtgg gtgttcggca acgccatgtg caagctgttc
     1561 accggcctgg cccacatcgg ctacttcggc ggcatcttct tcatcatcct gctgaccatc
     1621 gacagatacc tggccatcgt gcacgccgtg ttcgccctga aggccagaac cgtgaccttc
     1681 ggcgtggtga ccagcgtgat cacctggctg gtggccgtgt tcgccagcgt gcccggcatc
     1741 atcttcacca agtgccagaa ggaggacagc gtgtacgtgt gcggccccta cttccccaga
     1801 ggctggaaca acttccacac catcatgaga aacatcctgg gcctggtgct gcccctgctg
     1861 atcatggtga tctgctacag cggcatcctg aagaccctgc tgagatgcag aaacggcctg
     1921 gaggagggag gaaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac
     1981 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag
     2041 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
     2101 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2161 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     2221 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2281 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     2341 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2401 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2461 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     2521 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2581 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2641 aagctggagt acaacatcga caagaagaga cacagagccg tgagagtgat cttcaccatc
     2701 atgatcgtgt acttcctgtt ctggaccccc tacaacatcg tgatcctgct gaacaccttc
     2761 caggagttct tcggcctgag caactgcgag agcaccagcc agctggacca ggccacccag
     2821 gtgaccgaga ccctgggcat gacccactgc tgcatcaacc ccatcatcta cgccttcgtg
     2881 ggcgagaagt tcagaagata cctgagcgtg ttcttcagaa agcacatcac caagagattc
     2941 tgcaagcagt gccccgtgtt ctacagagag accgtggacg gcgtgaccag caccaacacc
     3001 cccagcaccg gcgagcagga ggtgagcgcc ggcctgatgg acgagctgta caagagtaga
     3061 atcacaagcg aaggcgagta catccccctg gatcaaatag acataaatgt aaagagtaga
     3121 atcacaagcg aaggcgagta catccccctg gatcaaatag acataaatgt aaagagtaga
     3181 atcacaagcg aaggcgagta catccccctg gatcaaatag acataaatgt aggtggattt
     3241 tgttatgaga atgaagtgta agatatcaag cttatcgata atcaacctct ggattacaaa
     3301 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3361 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3421 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3481 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3541 tgtcagctcc tttccggaac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3601 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3661 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3721 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3781 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3841 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     3901 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     3961 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     4021 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     4081 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     4141 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     4201 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     4261 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     4321 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4381 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4441 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4501 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4561 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4621 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4681 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4741 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4801 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4861 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4921 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4981 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5041 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5101 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5161 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5221 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5281 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5341 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5461 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5521 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6301 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7141 tttgctcaca tgt
//