LOCUS       sequence-356904-        3963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector backbone designed to express GFP from a EFS 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 3963)
  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 3963)
  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: "Mol 
            Brain."; date: "2015-02-24"; volume: "8:12. doi"; pages: " 
            10.1186/s13041-015-0100-7"
COMMENT     Sequence Label: pJEP223-AAV-EFS-GFP-pA
FEATURES             Location/Qualifiers
     source          1..3963
                     /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        170..381
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      405..414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(411..413,414..416,417..1130)
                     /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(411..413,414..416,417..1127)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(456..477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(717..736)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1064..1085
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1168..1187)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1226..1366
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1226..1355
                     /label=AAV2 ITR
     rep_origin      1441..1896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1528..1547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1738..1759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1913..1932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2032..2054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2092..2110)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2178..2282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2283..2351,2352..3143)
                     /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(2501..2520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3314..3902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3803..3822
                     /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 gcgttcgagt ggctccggtg cccgtcagtg ggcagagcgc
      181 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
      241 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
      301 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
      361 gggtttgccg ccagaacaca ggtgtcgtga cgcgggaatt cgccgccacc atggtgagca
      421 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      481 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      541 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      601 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      661 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      721 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      781 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      841 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
      901 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      961 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1021 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1081 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa ctcgagttta
     1141 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1201 ttttttcact ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     1261 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     1321 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     1381 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     1441 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1501 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1561 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1621 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     1681 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1741 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     1801 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     1861 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     1921 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1981 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2041 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2101 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2281 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2401 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2461 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2581 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2701 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2821 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2881 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3121 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3181 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3241 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3421 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3481 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3541 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3601 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3661 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3721 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3781 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3841 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3901 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3961 tgt
//
