LOCUS       sequence-12786-e        5185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5185)
  AUTHORS   Dittgen T, Nimmerjahn A, Komai S, Licznerski P, Waters J, Margrie 
            TW, Helmchen F, Denk W, Brecht M, Osten P
  TITLE     Lentivirus-based genetic manipulations of cortical neurons and their
            optical and electrophysiological monitoring in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Dec 28. 101(52):18206-11.
  PUBMED    15608064
REFERENCE   2  (bases 1 to 5185)
  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: "Proc Natl 
            Acad Sci U S A. 2004 Dec 28. 101(52):18206-11."
COMMENT     Sequence Label: AAV-CK(0.4)GW
FEATURES             Location/Qualifiers
     source          1..5185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(554..556,557..559,560..1273)
                     /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(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1314..1902
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1367..1387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1785..1796)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1905..1921)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1906..1922)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    1932..2408
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2131..2150)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2448..2588
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2448..2577
                     /label=AAV2 ITR
     rep_origin      2663..3118
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2750..2769)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2960..2981
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3135..3154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3254..3276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3314..3332)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3400..3504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3505..3573,3574..4365)
                     /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(3723..3742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4536..5124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5025..5044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaactt gtggactaag tttgttcaca
      181 tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg
      241 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa
      301 agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc
      361 ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca
      421 cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag
      481 gagcacgggc aggcgagtgg cccctagttc tgggggcagc tctagagccg atccccgggt
      541 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag taaagcggcc gcgactctag aattcgatat caagcttatc gataatcaac
     1321 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1381 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1441 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1501 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1561 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     1621 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1681 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     1741 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     1801 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     1861 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     1921 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     1981 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2041 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     2101 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     2161 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     2221 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     2281 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     2341 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     2401 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     2461 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2521 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     2581 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2641 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2701 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2761 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2821 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2881 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2941 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3001 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3061 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3121 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3181 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3241 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3301 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3361 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3421 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3481 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3541 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3601 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3661 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3721 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3781 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3841 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3901 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3961 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4021 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4081 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4141 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4201 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4261 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4321 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4381 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4441 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4501 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4561 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4621 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4681 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4741 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4801 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4861 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4921 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4981 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5041 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5101 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5161 ttttgctggc cttttgctca catgt
//
