LOCUS       Exported                7412 bp ds-DNA     circular SYN 26-AUG-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 16, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
FEATURES             Location/Qualifiers
     source          1..7412
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..141
                     /label=ITR-L
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    167..476
                     /label=mU6
     misc_feature    499..577
                     /label=Cj-sgRNA scaffold
     enhancer        683..986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        837..986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        987..1190
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    1215..1224
                     /label=Kozak sequence
     CDS             1227..1247
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1266..4463
                     /codon_start=1
                     /label=Hsp2Cas9
                     /translation="KILGFDIGIASVGWAYVEENELRDCGVRIFTKAENSTGESLALPR
                     REARGTRRRLKRRKQRLNELKRLLCKQFNFNLNDYCSNDGELAKAYQTTKDTKSPYELR
                     ALALEQKLDSKDLVRVILHIAKHRGYGNKHAKRDLEAEEKIKAKIEKEDQEITQKATQE
                     KQSTTTKGDSKKEQEQVLKALYRNETALKHYQTVGQYFYKEFKEKGQRIRNTTNNYQHT
                     MRQDWLKKELELIFEKQKEFGAKFSEGFENKIIEIAFFQRDLKSFEDKVGQCTFYPNEP
                     RAPKDSLSAIEFVTLTRIINTLKNLEEKSKNLGIGEVYGKDKIKEILNIVLDKGEISYK
                     KLREILGLNEQITFKDNKLDYAKDMKEEQKVKLIEFKNLKDFKKAIGGSFQAFGSFETF
                     DRKELDLIATDIAVIKSRENLAKKLENYPSLTQEQRESLSELNFSKHINLSLRALEKIL
                     PFMREGLRYNEAVEKAGLQEHRKHKQKSDFLPPLQDYEPYLANPVVERALSEYRKVLNA
                     LLRKYGQVHKIHLEFTREAKLSSKERQKYEKEQRENFARNQEAEKQCEAIGLPLNDAKN
                     ILKMKLWIEQRQLCAYSGEKITIEHLKDSTMLEIDHIYPYSRSYDDSYNNKVLCLAREN
                     QNKGNHTPFEAFGKDTEKWDRILSLASKLPKPKYRRISNKQFKDKEAGFLARNIVDTSY
                     IARLASEYTKHSLNFLPLSEDEVMITGEKDSKKHVEVVSGSLTATLRHYWGLGGKNGDK
                     DRDNHLHHVVDAIIIAYTNAKVIKAFSDFRKLQEQNRAKFYAKEIAQSKDKTKKGFEPP
                     CENFRNKIQEKVDSIFVSKPPRKRARGALHEETFYSLNDASLLKSYGGQEGVQRALELG
                     KIRRIGTKVVSNGEMKRVDIFKDKKGKFYGVPIYTMDFALGILPNKAVISGKKDGVIKD
                     WLEMDSNYEFCFSLFKDDLILVQKSEMEEPELCYYVGFDSCSPRIIVDKHNKCYKNMSD
                     NEKKLFTLKERERNNKKEEQKQDDENKEQKIIGGRLGIQNLKTFKKYQVSPLGEVKEVG
                     KETRQAIALKSTKHIKQD"
     CDS             4473..4493
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4494..4520
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4567..4648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             complement(4675..4815)
                     /label=ITR-R
     repeat_region   4675..4815
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4868..5174
                     /direction=RIGHT
                     /label=f1 origin
     rep_origin      4890..5345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5627..5731
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5681..6541
                     /label=Amp
     CDS             5732..6592
                     /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"
     rep_origin      6763..7351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcggcccgc tctagagatc cgacgcgcca
      181 tctctaggcc cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg ctactcccct
      241 gccccggtta atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa
      301 agacagataa tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct
      361 ataagagata caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct
      421 aactgtaaag taattgtgtg ttttgagact ataaatatcc cttggagaaa agccttgttt
      481 gcagtcttct cagaagacgt tttagtccct gaaaagggac taaaataaag agtttgcggg
      541 actctgcggg gttacaatcc cctaaaaccg cttttttact agtctcgagg gggggcccgg
      601 tacccaattc gccctatagt tctagttatt aatagtaatc aattacgggg tcattagttc
      661 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      721 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      781 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      841 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      901 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      961 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     1021 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     1081 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1141 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     1201 accgtcagat ccgcgccacc atggccccaa agaagaagcg gaaggtcggt atccacggag
     1261 tcccaaagat cctgggcttc gatatcggaa tcgccagcgt gggatgggcc tacgtggagg
     1321 aaaacgagct gagagattgc ggcgtgcgga tcttcacaaa ggccgagaat agcacaggtg
     1381 agtccctggc cctgcctcgg cgggaagcca gaggcacacg gagacgcctg aagcggagaa
     1441 aacagagact gaacgaactt aagagactgc tgtgtaaaca attcaacttc aacctgaacg
     1501 actactgcag caacgacggc gagctggcca aagcctatca gacaaccaag gacaccaaaa
     1561 gcccctatga gctgcgcgca ctggccctgg agcagaagct ggacagcaag gatctggtgc
     1621 gcgtcatcct gcacattgcc aagcacagag gctacggtaa caagcatgcc aaaagagacc
     1681 tggaagccga agaaaaaatc aaggccaaga tcgagaagga ggaccaggag atcacccaga
     1741 aagccaccca ggagaagcaa agcaccacca caaagggcga cagcaagaaa gagcaggagc
     1801 aggtgctgaa agctctgtac agaaatgaaa cagccctgaa acactaccag accgtgggcc
     1861 agtacttcta caaggagttc aaagagaaag gccaaagaat ccggaatacc accaacaact
     1921 accagcacac catgcggcag gactggctga agaaggaact ggagctgatc ttcgagaagc
     1981 agaaagagtt tggcgctaaa ttctctgaag gattcgagaa caagatcatc gagatcgcct
     2041 tcttccagag ggacctgaag tcttttgagg acaaggtggg ccaatgtacc ttctacccaa
     2101 acgaacctcg ggcccctaag gacagcctga gcgccatcga gttcgtgaca ctgacaagaa
     2161 tcatcaacac cctgaagaat ctggaggaaa agagcaagaa cctgggcata ggcgaagtgt
     2221 acggaaagga taagatcaag gagattctga acatcgtgct ggacaaaggc gagatcagct
     2281 acaagaagct gagggaaatc ctgggcctga acgagcagat cacattcaag gacaacaagc
     2341 tcgattacgc caaagacatg aaggaggaac aaaaagtgaa actgatcgag ttcaagaatc
     2401 tgaaagactt caaaaaggcc atcggcggaa gcttccaggc ttttggctcc tttgagacat
     2461 tcgacagaaa ggaactggat ctgatcgcaa ccgacatcgc cgtgatcaaa tccagagaaa
     2521 acctggctaa gaagctggaa aattatccca gcctgacaca ggagcaaaga gaatctctct
     2581 ctgagctcaa tttcagcaag cacatcaacc tgtctctgcg ggcccttgag aagatcctgc
     2641 cgttcatgag agagggcctg agatacaacg aggcagtcga gaaggccgga ctgcaggagc
     2701 acagaaagca caagcaaaag tccgacttcc tgcctcctct gcaggactac gagccatacc
     2761 tcgccaaccc cgtggtggaa agagccctga gcgaataccg gaaagttctg aacgccctgc
     2821 tgagaaagta cggccaggtg cacaagatcc acctggagtt caccagagag gctaagctga
     2881 gcagcaagga acggcagaag tacgagaagg agcaacggga aaacttcgcc agaaaccagg
     2941 aggccgagaa gcagtgcgag gccatcggcc tgcctctgaa tgatgccaaa aacatcctga
     3001 agatgaagct gtggattgag cagagacagc tgtgcgccta cagcggcgaa aagatcacca
     3061 ttgagcacct gaaggacagc acaatgctgg aaatcgacca catctaccct tacagcagat
     3121 cctacgatga ctcttacaac aacaaggtgc tgtgtctggc cagagaaaat cagaataaag
     3181 gcaaccacac accttttgag gccttcggca aggacacgga aaagtgggac cggatcctga
     3241 gcctcgctag caagctccct aaacctaagt acagaagaat cagcaacaag cagttcaagg
     3301 acaaggaggc cggcttcctg gctagaaaca tcgtggacac cagctatatc gcgagactgg
     3361 ccagtgaata cacaaagcat tctctgaact tcctgccact gtctgaggac gaggtgatga
     3421 tcaccggcga aaaggattct aaaaagcacg tcgaggtggt gagcggcagc ctcaccgcca
     3481 ccctgcggca ctactggggc ctgggcggaa agaacgggga caaggacaga gataaccacc
     3541 tgcaccacgt ggtggacgct atcatcatcg cttacaccaa cgccaaggtt atcaaggcct
     3601 tctctgattt caggaagctg caggagcaga acagagccaa gttctacgct aaggagatcg
     3661 cccagtctaa ggacaagacc aaaaaaggct ttgaacctcc atgtgaaaat ttccggaaca
     3721 agatccagga gaaggtcgac agtatcttcg tgagcaagcc cccccggaag agagccagag
     3781 gcgccctgca cgaggaaacc ttttatagcc tgaatgatgc cagccttctt aagtcttacg
     3841 gcggccagga gggcgtgcag agggcactgg agctgggcaa gatcagacgg atcggaacaa
     3901 aagtcgtgag caacggcgag atgaagagag tggatatctt caaggacaag aagggcaagt
     3961 tctacggagt gcccatctac accatggact tcgcgctggg aatcctgcct aacaaggccg
     4021 tgatcagcgg caagaaagac ggcgtgatca aggactggct ggaaatggac agcaactacg
     4081 aattctgctt ctctctgttt aaggatgacc tgatcctggt gcagaagtcc gagatggaag
     4141 agcccgagct gtgctactac gtgggctttg atagctgcag ccctcggatc atcgtggata
     4201 aacacaataa gtgctacaag aacatgagcg ataacgagaa gaaactgttc accctgaagg
     4261 aaagagagcg gaacaacaag aaggaggaac agaaacagga tgatgagaat aaggaacaga
     4321 agatcatcgg cggcagactc ggaattcaga acctgaaaac atttaagaag taccaagtgt
     4381 cccctctggg cgaggtgaag gaggtgggaa aagagaccag acaggccatc gccctgaaga
     4441 gcaccaagca tatcaagcag gactccggac ctccaaagaa aaagagaaaa gtatacccct
     4501 acgacgtgcc cgactacgcc taataactct cgagctcgat gagtttggac aaaccacaac
     4561 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4621 aaccattata agctgcaata aacaagttaa cgcacgtgcg gaccgagcgg ccgcaggaac
     4681 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4741 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4801 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4861 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4921 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4981 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5041 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5101 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5161 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5221 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5281 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5341 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5401 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5461 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5521 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6121 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6241 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6901 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7141 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7381 cctggccttt tgctggcctt ttgctcacat gt
//
