LOCUS       sequence-370039-        5202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging vector that expresses secreted C-CDNF under control
            of the EGFP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5202)
  TITLE     Harvey Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5202)
  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"
COMMENT     Sequence Label: pOTTC1584 - pscAAV CMV-IE secreted EGFP-THPKTE...
FEATURES             Location/Qualifiers
     source          1..5202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        236..539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        540..743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     694..714
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(868..870,871..873,874..1584)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(868..870,871..873,874..1584)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(913..934)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1174..1193)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1521..1542
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1649..2237
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1702..1722)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2120..2131)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2279..2296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2285..2396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2508..2648
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2508..2637
                     /label=AAV2 ITR
     rep_origin      2723..3178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2810..2829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3020..3041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3195..3214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3314..3336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3374..3392)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3460..3564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3565..3633,3634..4425)
                     /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(3783..3802)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4596..5184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5085..5104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc tgcggccgca cgcgtggatc taatatagtt
      181 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      721 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgaagt tatcgctagc
      781 accatgtggt gcgcgagccc agttgctgtg gtggcctttt gcgccgggct tttggtctct
      841 cacccggtgc tgacgcaggg ccaggagatg gtgagcaagg gcgaggagct gttcaccggg
      901 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      961 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1021 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     1081 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1141 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1201 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1261 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1321 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1381 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1441 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1501 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1561 ctcggcatgg acgagctgta caagacacac cccaaaacag agctctaggg cgcgccataa
     1621 cttcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     1681 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1741 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1801 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     1861 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     1921 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     1981 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2041 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     2101 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2161 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2221 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctaatt cctagaagcc
     2281 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2341 accctggaag gtgccactcc caccgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2401 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2461 gattgggaag acaatagcag gcatgctggg gcggaccgag cggccgcagg aacccctagt
     2521 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2581 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     2641 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2701 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2761 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     2821 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2881 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2941 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3001 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     3061 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     3121 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3181 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3241 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3301 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3361 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3421 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3481 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3541 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3601 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3661 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3721 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3781 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3841 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3901 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3961 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4021 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4081 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4141 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4201 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4261 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4321 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4381 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4441 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4501 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4561 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4621 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4681 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4741 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4801 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4861 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4921 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4981 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5041 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5101 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5161 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cc
//
