LOCUS       sequence-321943-        7313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express SV-Tag construct in a Cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7313)
  AUTHORS   Chantranupong L, Saulnier JL, Wang W, Jones DR, Pacold ME, Sabatini 
            BL
  TITLE     Rapid purification and metabolomic profiling of synaptic vesicles 
            from mammalian brain.
  JOURNAL   Elife. 2020 Oct 12;9. pii: 59699. doi: 10.7554/eLife.59699.
  PUBMED    33043885
REFERENCE   2  (bases 1 to 7313)
  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: "Elife."; 
            date: "2020-10-12"; volume: "9. pii: 59699. doi"; pages: " 
            10.7554/eLife.59699"
COMMENT     Sequence Label: pAAV hSynapsin flexed SV-tag
FEATURES             Location/Qualifiers
     source          1..7313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2443..2572
                     /label=AAV2 ITR
     polyA_signal    2734..2855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2771..2790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2825..2844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2884..2900
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2885..2901
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2904..3492
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3010..3021
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3419..3439
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(3505..3538)
                     /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(3589..3622)
                     /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)."
     regulatory      3641..3650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3780..3798)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4048..4067
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4242..4265
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             5081..5134
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6095..6121
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6134..6160
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6173..6199
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6212..6238
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6251..6277
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6290..6316
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6329..6355
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6368..6394
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6407..6433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    6451..6484
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6535..6568
                     /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)."
     promoter        complement(6584..7031)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   7113..7242
                     /label=AAV2 ITR
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2401 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctgcgcgc tcgctcgctc
     2461 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     2521 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta
     2581 atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagat ccttatcgat
     2641 tttaccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacatct ccccctgaac
     2701 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2761 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2821 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa gcggccgccc
     2881 gggtcgaggt cgacggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     2941 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     3001 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     3061 ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     3121 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     3181 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
     3241 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     3301 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     3361 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     3421 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     3481 cagaggttga ttatcgataa gcttataact tcgtatagca tacattatac gaagttattc
     3541 tttgcaccat tctaaagaat aacagtgata atttctgggt taaggcaaat aacttcgtat
     3601 aggatacttt atacgaagtt atctcgagac tagtaccggt gccaccatgg tgagcaaggg
     3661 cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     3721 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3781 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     3841 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     3901 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     3961 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     4021 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4081 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     4141 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     4201 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     4261 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     4321 cctggggcat ggtaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     4381 caacaacatg gccgtgatta aggagtttat gaggtttaaa gtcaggatgg agggcagcat
     4441 gaacgggcac gagtttgaga ttgaaggaga gggggaaggg aggccctatg agggcaccca
     4501 gacagcaaag ctgaaagtga caaagggcgg ccctctgcca ttcgcatggg acatcctgtc
     4561 tcctcagttt atgtatggaa gtaaagccta cgtgaaacac cctgctgaca ttccagatta
     4621 caagaaactg agcttcccag aggggtttaa gtgggaaaga gtgatgaact tcgaagacgg
     4681 gggcctggtg actgtcaccc aggacagctc tctgcaggac ggcactctga tctataaggt
     4741 gaaaatgcgc ggcaccaatt ttccccctga tggcccagtc atgcagaaga agacaatggg
     4801 ctgggaggct tcaacagaaa ggctgtaccc cagagacggc gtgctgaagg gagagatcca
     4861 ccaggcactg aagctgaaag atggagggca ttatctggtc gagttcaaaa ccatctacat
     4921 ggccaagaaa cccgtgcagc tgcctggcta ctattacgtc gacacaaagc tggatattac
     4981 tagccataac gaagactaca ccattgtgga gcagtatgag cggagcgagg ggaggcacca
     5041 tctgtttctg tacggcatgg acgagctgta caagactagt gagggcagag gaagtcttct
     5101 gacatgcgga gacgtggaag agaatcccgg ccctgcccca ggctcatcta gaatggacgt
     5161 ggtgaatcag ctggtggctg ggggtcagtt ccgggtggtc aaggagcccc ttggcttcgt
     5221 gaaggtgctg cagtgggtct ttgccatctt cgcctttgct acgtgcggca gctacaccgg
     5281 agagcttcgg ctgagcgtgg agtgtgccaa caagacggag agtgccctca acatcgaagt
     5341 cgaatttgag tacccattca ggctgcacca agtgtacttt gatgcaccct cctgcgttaa
     5401 agggggcact accaagatct tcctagttgg tgactactcc tcctcggctg agttctttgt
     5461 caccgtggct gtgtttgcct tcctctactc catgggggcc ctggccacct acatcttcct
     5521 gcagaacaag taccgagaga acaacaaagg gccaatgatg gacttcctgg ccacagcagt
     5581 gttcgctttc atgtggctag ttagctcatc cgcctgggcc aaaggcctgt ccgatgtgaa
     5641 gatggccact gacccagaga acattatcaa ggagatgcct atgtgccgcc agacaggaaa
     5701 cacatgcaag gaactgaggg accctgtgac ttcaggactc aacacctcgg tggtgtttgg
     5761 cttcctgaac ctggtgctct gggttggcaa cctatggttc gtgttcaagg agacaggctg
     5821 ggccgcccca ttcatgcgcg cacctccagg cgccccagaa aagcaaccag ctcctggcga
     5881 tgcctacggc gatgcgggct atgggcaggg ccccggaggc tatgggcccc aggactccta
     5941 cgggcctcag ggtggttatc aacccgatta cgggcagcca gccagcggcg gtggcggtgg
     6001 ctacgggcct cagggcgact atgggcagca aggctacggc caacagggtg cgcccacctc
     6061 cttctccaat cagatgtcgc gaggagggag cggctatccc tatgacgtgc ctgattacgc
     6121 cggcacagga tcctacccct atgatgtgcc tgactacgct ggcagcgccg gataccctta
     6181 tgatgtgcct gattatgctg gagggagcgg ctacccatac gatgttccag attacgctgg
     6241 aacaggcagc tatccctatg acgtcccgga ctatgcaggc agcgccggat atccatatga
     6301 cgttccagat tacgctggag gctccggcta cccttatgat gtgcccgatt atgccggaac
     6361 aggcagctat ccctatgacg tgcctgacta tgctggatcc gccggctacc cttatgacgt
     6421 gcccgattac gcctaagcgg ccgcactagt ataacttcgt ataatgtatg ctatacgaag
     6481 ttattaagag gtttcatatt gctaatagca gctacaatcc agctaccatt ctgcataact
     6541 tcgtataaag tatcctatac gaagttataa gatctgaatt cgactgcgct ctcaggcacg
     6601 acacgactcc tccgctgccc accgcagact gaggcagcgc tgagtcgccg gcgccgcagc
     6661 gcagatggtc gcgcccgtgc ccccctatct cgcgcctcgc gtggtgcggt ccggctgggc
     6721 cggcggcggc gcggacgcga ccaaggtggc cgggaagggg agtttgcggg ggaccggcga
     6781 gtgacgtcag cgcgccttca gtgctgaggc ggcggtggcg cgcgccgcca ggcgggggcg
     6841 aaggcactgt ccgcggtgct gaagctggca gtgcgcacgc gcctcgccgc atcctgtttc
     6901 ccctccccct ctctgatagg ggatgcgcaa tttggggaat gggggttggg tgcttgtcca
     6961 gtgggtcggg gtcggtcgtc aggtaggcac ccccaccccg cctcatcctg gtcctaaaac
     7021 ccacttgcac tcatacgcag ggccctctgc agagatcttc ctagagcatg gctacgtaga
     7081 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     7141 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     7201 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctggcgta atagcgaaga
     7261 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggg
//
