LOCUS       DRH001_scAAV2-hS        5963 bp DNA     circular SYN 11-APR-2023
DEFINITION  from NGVB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1793..2244
                     /mol_type="other DNA"
                     /db_xref="taxon:1476495"
                     /organism="Cloning vector pAAV-CW3SL-EGFP"
     LTR             7..112
                     /label=AAV2 Mut ITR
                     /label=AAV2\Mut\ITR; AAV2\Mut\ITR (delta TRS)
     promoter        174..621
                     /label=hSyn promoter
     primer_bind     184..203
                     /label=P023_hSyn_F
     primer_bind     complement(280..298)
                     /label=P018_AAV_hSyn_through5primeITR_R
     misc_feature    635..644
                     /label=kozak
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             650..670
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             674..694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             698..718
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(699..718)
                     /label=P048_Sv40NLS_R
     misc_feature    719..1744
                     /locus_tag="iCre"
                     /label=iCre
     primer_bind     complement(771..790)
                     /label=P025_iCre_R
     primer_bind     1003..1022
                     /label=P056_iCre_F
     primer_bind     complement(1416..1435)
                     /label=P032_iCre_2_R
     primer_bind     1659..1678
                     /label=p055_iCre_3primeEnd_F
     CDS             1760..1786
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1809..2059
                     /label=WPRE3
     primer_bind     complement(1885..1904)
                     /label=P024_WPRE3_R
     primer_bind     2039..2057
                     /label=P019_WPRE3_through3primeITR_F
     misc_feature    2060..2240
                     /label=SV40 Late poly(A) signal sequence
     primer_bind     complement(2220..2238)
                     /label=P050_SV40pA_through3primeITR_F
     LTR             2245..2414
                     /label=AAV2 ITR
                     /label=AAV2\ITR
     primer_bind     2453..2467
                     /label=P007_scAAV_F1Ori
     primer_bind     complement(2482..2501)
                     /label=P006_scAAV_ITR_3prime_Rev
     primer_bind     2498..2517
                     /label=P026_scAAV_F1Ori_v2
     primer_bind     complement(2546..2565)
                     /label=P020_scAAV_ITR_3prime_v2_R
     rep_origin      2746..3259
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2746..3201
                     /direction=RIGHT
                     /label=f1 ori
     primer_bind     complement(2833..2852)
                     /label=F1ori-R
     primer_bind     complement(2833..2852)
                     /label=P015_AAV_F1Ori_through3primeITR_R
     primer_bind     complement(2833..2852)
                     /label=P015_AAV_F1Ori_through3primeITR_Ra
     primer_bind     2835..2854
                     /label=P031_scAAV_F1Ori_F
     primer_bind     complement(2943..2961)
                     /label=P057_AAV_F1Ori_through3primeITR_2_R
     primer_bind     2963..2982
                     /label=P008_scAAV_F1Ori_2
     primer_bind     3043..3064
                     /label=F1ori-F
     primer_bind     3419..3438
                     /label=P027_scAAV_AmpR_v2
     primer_bind     3432..3451
                     /label=P009_scAAV_AmpR
     primer_bind     complement(3741..3760)
                     /label=pRS-marker
     primer_bind     3860..3882
                     /label=pGEX 3'
     primer_bind     3914..3933
                     /label=P010_scAAV_AmpR2
     primer_bind     complement(3920..3938)
                     /label=pBRforEco
     primer_bind     3966..3985
                     /label=P028_scAAV_AmpR_2_v2
     promoter        4006..4110
                     /gene="bla"
                     /label=bla promoter
                     /label=AmpR promoter
     CDS             4111..4971
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /label=AmpR
     primer_bind     complement(4329..4348)
                     /label=Amp-R
     primer_bind     4345..4365
                     /label=P011_scAAV_AmpR3
     primer_bind     4782..4801
                     /label=P029_scAAV_AmpR_3_v2
     primer_bind     4901..4922
                     /label=P012_scAAV_puc1
     rep_origin      5119..5786
                     /direction=RIGHT
                     /label=pUC ori
     rep_origin      5142..5730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5358..5377
                     /label=P030_scAAV_puc_v2
     primer_bind     5383..5402
                     /label=P013_scAAV_puc2
     primer_bind     5509..5528
                     /label=P016_AAV_puc3_through5primeITR_F
     primer_bind     5631..5650
                     /label=pBR322ori-F
     primer_bind     5787..5807
                     /label=P021_scAAV_ITR_5prime_v2_F
     primer_bind     5841..5860
                     /label=P000_scAAV-ITR
     primer_bind     5884..5901
                     /label=L4440
     misc_feature    5920..5963
                     /label=added the 50bp leading up to mutated ITR
ORIGIN
        1 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggggttcggt
      121 accgcggccg cacgcgttgt acagtgtcta gactgcagag ggccctgcgt atgagtgcaa
      181 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
      241 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      301 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      361 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      421 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      481 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      541 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      601 gtgtcgtgcc tgagagcgca gtcgagaagg atccgccacc atgggtggac caaaaaagaa
      661 gagaaaggta gacccgaaga aaaagcgtaa ggtcgatccc aagaagaaga ggaaggtgtc
      721 caacctgctg actgtgcacc aaaacctgcc tgccctccct gtggatgcca cctctgatga
      781 agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg
      841 gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acaacaggaa
      901 atggttccct gctgaacctg aggatgtgag ggactacctc ctgtacctgc aagccagagg
      961 cctggctgtg aagaccatcc aacagcacct gggccagctc aacatgctgc acaggagatc
     1021 tggcctgcct cgcccttctg actccaatgc tgtgtccctg gtgatgagga gaatcagaaa
     1081 ggagaatgtg gatgctgggg agagagccaa gcaggccctg gcctttgaac gcactgactt
     1141 tgaccaagtc agatccctga tggagaactc tgacagatgc caggacatca ggaacctggc
     1201 cttcctgggc attgcctaca acaccctgct gcgcattgcc gaaattgcca gaatcagagt
     1261 gaaggacatc tcccgcaccg atggtgggag aatgctgatc cacattggca ggaccaagac
     1321 cctggtgtcc acagctggtg tggagaaggc cctgtccctg ggggttacca agctggtgga
     1381 gagatggatc tctgtgtctg gtgtggctga tgaccccaac aactacctgt tctgccgggt
     1441 cagaaagaat ggtgtggctg ccccttctgc cacctcccaa ctgtccaccc gggccctgga
     1501 agggatcttt gaggccaccc accgcctgat ctatggtgcc aaggatgact ctgggcagag
     1561 atacctggcc tggtctggcc actctgccag agtgggtgct gccagggaca tggccagggc
     1621 tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg accaatgtga acattgtgat
     1681 gaactacatc agaaacctgg actctgagac tggggccatg gtgaggctgc tcgaggatgg
     1741 ggacggtgga ggcggaagtt acccatacga tgttccagat tacgcttaat gaatcgaatt
     1801 ccgctcgaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1861 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1921 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     1981 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2041 ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2101 tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2161 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2221 tgggaggttt tttaaagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2281 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2341 cccgggcggc ctcagtgagc gagcgagcgc gccagctggc gtaatagcga agaggcccgc
     2401 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgatt ccgttgcaat
     2461 ggctggcggt aatattgttc tggatattac cagcaaggcc gatagtttga gttcttctac
     2521 tcaggcaagt gatgttatta ctaatcaaag aagtattgcg acaacggtta atttgcgtga
     2581 tggacagact cttttactcg gtggcctcac tgattataaa aacacttctc aggattctgg
     2641 cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg tttagctccc gctctgattc
     2701 taacgaggaa agcacgttat acgtgctcgt caaagcaacc atagtacgcg ccctgtagcg
     2761 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2821 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2881 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2941 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3001 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3061 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3121 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3181 aaatattaac gcttacaatt taaatatttg cttatacaat cttcctgttt ttggggcttt
     3241 tctgattatc aaccggggta catatgattg acatgctagt tttacgatta ccgttcatcg
     3301 attctcttgt ttgctccaga ctctcaggca atgacctgat agcctttgta gagacctctc
     3361 aaaaatagct accctctccg gcatgaattt atcagctaga acggttgaat atcatattga
     3421 tggtgatttg actgtctccg gcctttctca cccgtttgaa tctttaccta cacattactc
     3481 aggcattgca tttaaaatat atgagggttc taaaaatttt tatccttgcg ttgaaataaa
     3541 ggcttctccc gcaaaagtat tacagggtca taatgttttt ggtacaaccg atttagcttt
     3601 atgctctgag gctttattgc ttaattttgc taattctttg ccttgcctgt atgatttatt
     3661 ggatgttgga atcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3721 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3781 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3841 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3901 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3961 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4021 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4081 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4141 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4201 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4261 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4321 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4381 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4441 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4501 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4561 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4621 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4681 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4741 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4801 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4861 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4921 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4981 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5041 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5101 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5161 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5221 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5281 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5341 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5401 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5461 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5521 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5581 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5641 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5701 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5761 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5821 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5881 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5941 gcgcgttggc cgattcatta atg
//
