LOCUS       sequence-356881-        4824 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV vector backbone designed to express GFP from a 1.1Synapsin 
            promoter. It also contains a poly-Adenylation Signal (pA) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4824)
  AUTHORS   Holehonnur R, Lella SK, Ho A, Luong JA, Ploski JE
  TITLE     The production of viral vectors designed to express large and 
            difficult to express transgenes within neurons.
  JOURNAL   Mol Brain. 2015 Feb 24;8:12. doi: 10.1186/s13041-015-0100-7.
  PUBMED    25887710
REFERENCE   2  (bases 1 to 4824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Brain."; date: "2015-02-24"; volume: "8:12. doi"; pages: " 
            10.1186/s13041-015-0100-7"
COMMENT     Sequence Label: pJEP219-AAV-1.1Synapsin-GFP-pA
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1266..1275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1272..1274,1275..1277,1278..1991)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1272..1274,1275..1277,1278..1988)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1317..1338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1578..1597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1925..1946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2029..2048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2087..2227
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2087..2216
                     /label=AAV2 ITR
     rep_origin      2302..2757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2389..2408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2599..2620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2774..2793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2893..2915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2953..2971)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3039..3143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3144..3212,3213..4004)
                     /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(3362..3381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4175..4763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4664..4683
                     /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 gcgtgggttt tggctacgtc cagagcagag gaatgagggc
      181 atgtagacta aatatgttcg tgtggaagag gctgaataca catcagagtt agtgctgcag
      241 gaaatgcttc tgcattgcat acccagagtt tccttgctca tctgagagca tgtgtttttt
      301 ccagatgtgt gtacttgtgt gagattctct gggtgtgtgt caatgtgttg cctgaacgtg
      361 cattgctcaa tatgctcatg tgtgttaccc tgggcttgta catctacata tatacctgga
      421 tgcccgtgtg ttctgtgatg tacatatacc ctgtgtcatt ccttgttttt ctatttgtgt
      481 tattccatgt gttccttcag gctctcacta cccaagtgtc cacctccgcc tgtctggtga
      541 tgtttacgct accccgtgct cttttctttg cctgacagtg ttgtcgtgga agacatctcg
      601 ccaggaacac tgcagtaagg agaatttcta gttttatgtt cccctccgag tatgcttcta
      661 tcccgaccct caaccccaaa atgccttcag aggtgaaaat caacactgga aacacaagta
      721 tctgggaagg gtaacaatgc aagttagcct gaggatttag gaggaggctg aaaaacagag
      781 taggagcctt actacgggtc cagaccctac ggacaagaac ccccactccc actccccaaa
      841 ttgcgcattc cctcccccat cagaggggga ggggaagagg atgcagcgcg gcgcggcgcg
      901 tgcgcactgt cggatttagt accgcggaca gagccttcgc ccccgctgcc ggcgcgcgcc
      961 accacctccc cagcaccaaa ggcgggctga cgtcactctc cagccctccc caaactcccc
     1021 tacctcaccg ccttggtcgc gtccgtgcag cggtgagtcc agtcgggccg caccacaaga
     1081 ggtgcaagat aggggggtgc aggcgcgacc atacgctctg cggcggcaga gcctcagcgc
     1141 tgcctcagtc tgcagcgggc agcagaggag tcgcgtcgtg ccagagagcg ccgccgtgct
     1201 cctgagcccc ttgcgctccg cccccgcggc ccaccgaccc actgcccctt ggatccgaat
     1261 tcgccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1321 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1381 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1441 cctggcccac tctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1501 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1561 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1621 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1681 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1741 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1801 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1861 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1921 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1981 tgtacaagta actcgagttt attgcagctt ataatggtta caaataaagc aatagcatca
     2041 caaatttcac aaataaagca tttttttcac tggaccgagc ggccgcagga acccctagtg
     2101 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2161 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2221 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2281 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2341 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2401 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2461 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2521 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2581 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     2641 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     2701 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     2761 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2821 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2881 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2941 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3001 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4801 tttgctggcc ttttgctcac atgt
//
