LOCUS       sequence-436147-        5777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Cre-dependent enriched expression of GCaMP8f in axons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5777)
  TITLE     For Cre-dependent enriched expression of GCaMP8f in axons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5777)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-syn-DIO-axon-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..5777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1351..1370
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1610..1631
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1737..1758)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2040..2057)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2124..2157
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2208..2241
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2266..2854
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2319..2339)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2737..2748)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2908..2927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2962..2981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3040..3180
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3040..3169
                     /label=AAV2 ITR
     rep_origin      3255..3710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3342..3361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3552..3573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3727..3746)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3846..3868
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3906..3924)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3992..4096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4097..4165,4166..4957)
                     /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(4315..4334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5128..5716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5617..5636
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgcctccgg atcccttcgc tgtcatcatt tgtacaaact cttcgtagtt tacctgacca
      841 tccccatcga tgtctgcttc cctgatcatt tcatcaacct cttcatctgt taacttctct
      901 ccaaggtttg tcatcacgtg gcgaagctct gctgcactga tgtagccatt gccatcctta
      961 tcaaacacac cgaacgcttc tctaatttct tcttccgtgt ccctgtattt catttttctt
     1021 gccatcattg tcaggaactc agggaagtcg attgtgccgt caccgtcggc atctacttca
     1081 ttgatcatgt cctgcagctc tgcttctgtg gggttcagtc cgagagaccg catcaccgtc
     1141 cccagctcct tggttgttat tgtcccatcc ccgtccttgt caaataggga gaaagcctct
     1201 ttaaattctg cgatctgctc ttcagtcagt tggtccggca ggttgtactc cagcttgtgc
     1261 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1321 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1381 atggtgcgct cctggatgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1441 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     1501 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     1561 gtggcatcgc cctcaccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     1621 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatgctc
     1681 cctccggtac cgcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     1741 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttcgaaag tttggactcc
     1801 acgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     1861 tggtagtggt aggcgagctg cacgccgccg tcctcgatgt tgtggcggat gtggaagttc
     1921 gccttgatgc cgttcttctg cttgtcggcc ttgatataga cgttgatctt caggcccatc
     1981 aggcgggcga tgatcttcac ggcggtggcc acctccttga aggtcttctt gcgacgcgtg
     2041 tgatgatggt gatgatgaat cttttggtcc tcatcattct tttcaacctg tttggttctt
     2101 ctcatacagc acagcatgct agcataactt cgtataaagt atcctatacg aagttatttg
     2161 ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata acttcgtata
     2221 atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca acctctggat
     2281 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2341 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2401 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2461 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2521 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2581 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2641 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2701 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2761 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2821 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg
     2881 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2941 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3001 agctgcaata aactaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3061 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3121 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3181 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3241 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3301 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     3361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     3481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3541 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     3601 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     3661 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     3721 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3781 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3841 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3901 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3961 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4021 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4081 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4141 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4201 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4261 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4321 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4381 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4441 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4501 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4561 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4621 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4681 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4741 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4801 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4861 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4921 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4981 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5041 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5101 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5161 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5221 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5281 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5341 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5401 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5461 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5521 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5581 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5641 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5701 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5761 gccttttgct cacatgt
//
