LOCUS       sequence-10978-e        6106 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 6106)
  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 6106)
  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(1.3)GW
FEATURES             Location/Qualifiers
     source          1..6106
                     /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
     promoter        158..1447
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1469..1478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1475..1477,1478..1480,1481..2194)
                     /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(1475..1477,1478..1480,1481..2191)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1520..1541)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1781..1800)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2128..2149
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2235..2823
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2288..2308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2706..2717)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2826..2842)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2827..2843)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2853..3329
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3369..3509
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3369..3498
                     /label=AAV2 ITR
     rep_origin      3584..4039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3881..3902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4056..4075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4175..4197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4235..4253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4321..4425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4426..4494,4495..5286)
                     /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(4644..4663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5457..6045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5946..5965
                     /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 cgcgtaacat tatggcctta ggtcacttca
      181 tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca gagagcttcc
      241 tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa ctcagattac
      301 taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg agtgaactca
      361 ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta catctacacc
      421 acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc ctctctccag
      481 tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga gaccaaatct
      541 tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca ggaggggccc
      601 tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc gtcagcattc
      661 tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca caatggctaa
      721 aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca accagaccaa
      781 ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga aggagagatg
      841 aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg ggcaatacct
      901 gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct caaggaccat
      961 cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc ccacttaatc
     1021 ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga agagcgatga
     1081 tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta catcaccctg
     1141 ggagaacaag gtccacttgc ttctgggccc acacagtcct gcagtattgt gtatataagg
     1201 ccagggcaaa ggaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt
     1261 accggggcaa cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac
     1321 gaaggctcgc gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag
     1381 tcaagccggt tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg
     1441 gggcagcggg gatccccggg taccggtcgc caccatggtg agcaagggcg aggagctgtt
     1501 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1561 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1621 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1681 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1741 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1801 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1861 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1921 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1981 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2041 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2101 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2161 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gaattcgata
     2221 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2281 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2341 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2401 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2461 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2521 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2581 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2641 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2701 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2761 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2821 cgcatcgata ccgtcgacct cgagagatct acgggtggca tccctgtgac ccctccccag
     2881 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     2941 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3001 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3061 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3121 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3181 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3241 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3301 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3361 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3421 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3481 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3541 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3661 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3781 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3901 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3961 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4021 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4261 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5581 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
