LOCUS       sequence-334101-        6278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent HaloTag labeling of neuronal presynaptic terminals 
            from the synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6278)
  TITLE     AAV constructs for HaloTag labeling of neuronal presynaptic 
            terminals
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6278)
  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            "Unpublished"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-phSyn-flex-SypHaloTag
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(481..514)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(525..558)
                     /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)."
     CDS             complement(572..1462)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     regulatory      2389..2398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2416..2449
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2460..2493
                     /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)."
     misc_feature    2517..3105
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2570..2590)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2988..2999)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3140..3157)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3146..3353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3376..3516
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3376..3505
                     /label=AAV2 ITR
     rep_origin      3591..4046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3888..3909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4063..4082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4182..4204
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4242..4260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4328..4432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4433..4501,4502..5293)
                     /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(4651..4670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5464..6052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5953..5972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6114..6243
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagac gcgtggacgg tacccttcag gtcgacgtcg
      481 ataacttcgt atagcataca ttatacgaag ttatccgaat cgcaataact tcgtatagga
      541 tactttatac gaagttattt ggcgcgcctt agccggaaat ctcgagcgtc gacagccagc
      601 gcgcgatctc gctgccgatc aggtccgggt tgtcttcttg cagcagattc agaccagggc
      661 cgatgtccac agccttgcag ttaggcaggc ttttggccag gcgagcggct tcggccggtg
      721 ggatcagaac gcctggggtg ccccagaaca gcagcttcgg gacaggggac tggtgcagcc
      781 agtccatgta ttcttcgacc agcgcgacga tgttcgctgg ctcaccggcg attggcagct
      841 cgtttgggaa gcgccacagt ggctcgcggt caacaggatt caggaacggc tcgcggtaat
      901 ggtccatctc gacttcagtc agcgggcgga cgacacccat cggcagcgta ccctcgataa
      961 aaacgttctg atcgatgatc agcttgcggc cgacgtcggt ggtgcggaag gcctggaagg
     1021 tctcgcgggc aaattctggc cattcgtccc aggtcgggat agggcggatg aactccataa
     1081 atgcaatacc tttgacgcgc tctggattgc gcttggccca gtggaaaccc agagcggagc
     1141 cccagtcgtg aatgaccagg acgacctctt ccagacccag ggcttcgatg aaggcatcca
     1201 tgaagcggac gtggtcgtcg aagaaataac ccaggtctgg tttgtcggat ttgcccatac
     1261 cgatcaggtc tggagcaatg cagcgatggg tcggtgcaac atgcgggatg atgttgcgcc
     1321 acacgtagga ggaggtcggg ttaccgtgca ggaacagcac aggggtgcca tcgcgcggac
     1381 caacatcgac gtagtgcatg cgctcgccca ggacttccac ataatggggg tcgaatggaa
     1441 agccagtacc gatttctgcc attcgcgaca tctgattgga gaaggaggtg ggcgcaccct
     1501 gttggccgta gccttgctgc ccatagtcgc cctgaggccc gtagccaccg ccaccgccgc
     1561 tggctggctg cccgtaatcg ggttgataac caccctgagg cccgtaggag tcctggggcc
     1621 catagcctcc ggggccctgc ccatagcccg catcgccgta ggcatcgcca ggagctggtt
     1681 gcttttctgg ggcgcctgga ggtgcgcgca tgaatggggc ggcccagcct gtctccttga
     1741 acacgaacca taggttgcca acccagagca ccaggttcag gaagccaaac accaccgagg
     1801 tgttgagtcc tgaagtcaca gggtccctca gttccttgca tgtgtttcct gtctggcggc
     1861 acataggcat ctccttgata atgttctctg ggtcagtggc catcttcaca tcggacaggc
     1921 ctttggccca ggcggatgag ctaactagcc acatgaaagc gaacactgct gtggccagga
     1981 agtccatcat tggccctttg ttgttctctc ggtacttgtt ctgcaggaag atgtaggtgg
     2041 ccagggcccc catggagtag aggaaggcaa acacagccac ggtgacaaag aattcagccg
     2101 aggaggagta gtcaccaact aggaagatct tggtagtgcc ccctttaacg caggagggtg
     2161 catcaaagta cacttggtgc agcctgaatg ggtactcaaa ttcgacttcg atgttgaggg
     2221 cactctccgt cttgttggca cactccacgc tcagccgaag ctctccggtg tagctgccgc
     2281 acgtagcaaa ggcgaagatg gcaaagaccc actgcagcac cttcacgaag ccaaggggct
     2341 ccttgaccac ccggaactga cccccagcca ccagctgatt caccacgtcc atggtggcca
     2401 acctggccgg ccttgataac ttcgtataat gtatgctata cgaagttatc cgaatcgcaa
     2461 taacttcgta taaagtatcc tatacgaagt tatcgaagat catcaagctt atcgataatc
     2521 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2581 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2641 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2701 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2761 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2821 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2881 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2941 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3001 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3061 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3121 tgctcgagag atcgatctgc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3181 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3241 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3301 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggacacgtgc
     3361 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3421 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3481 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3541 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3661 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3781 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3901 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3961 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4021 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4081 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4141 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4201 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4261 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4321 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4381 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4441 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4501 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4561 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4621 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4681 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4741 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4801 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4861 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4921 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4981 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5041 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5101 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5161 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5221 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5281 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5341 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5401 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5461 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5521 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5581 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5641 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5701 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5761 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5821 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5881 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5941 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6001 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6061 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     6121 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     6181 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     6241 cctgcggccg cacgcgtctg cagagggccc tgcgtatg
//
