LOCUS       V014724                 8824 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014724
VERSION     V014724
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8824)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             837..4736
                     /gene="cas12a"
                     /label="CRISPR-associated endonuclease Cas12a"
                     /note="CRISPR-associated endonuclease Cas12a from
                     Francisella tularensis subsp. novicida (strain U112).
                     Accession#: A0Q7Q2"
     CDS             4770..5486
                     /label="EGFP"
                     /note="enhanced GFP"
     polyA_signal    5612..5733
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5740..6195)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6222..6326
                     /label="AmpR promoter"
     promoter        6328..6685
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             6720..7511
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    7746..7793
                     /label="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8122..8710
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatggcct ccactcgtgt cctcgcctct cgcctggcct
      661 cccggatggc tgcttccgcc aaggttgccc gccctgctgt ccgcgttgct caggtcagca
      721 agcgcaccat ccagactggc tcccccctcc agaccctcaa gcgcacccag atgacctcca
      781 tcgtcaacgc caccacccgc caggctttcc agaagcgcgc ctactcttcc ggttctatga
      841 gcatctacca ggagttcgtc aacaagtatt cactgagtaa gacactgcgg ttcgagctga
      901 tcccacaggg caagacactg gagaacatca aggcccgagg cctgattctg gacgatgaga
      961 agcgggcaaa agactataag aaagccaagc agatcattga taaataccac cagttcttta
     1021 tcgaggaaat tctgagctcc gtgtgcatca gtgaggatct gctgcagaat tactcagacg
     1081 tgtacttcaa gctgaagaag agcgacgatg acaacctgca gaaggacttc aagtccgcca
     1141 aggacaccat caagaaacag attagcgagt acatcaagga ctccgaaaag tttaaaaatc
     1201 tgttcaacca gaatctgatc gatgctaaga aaggccagga gtccgacctg atcctgtggc
     1261 tgaaacagtc taaggacaat gggattgaac tgttcaaggc taactccgat atcactgata
     1321 ttgacgaggc actggaaatc atcaagagct tcaagggatg gaccacatac tttaaaggct
     1381 tccacgagaa ccgcaagaac gtgtactcca gcaacgacat tcctacctcc atcatctacc
     1441 gaatcgtcga tgacaatctg ccaaagttcc tggagaacaa ggccaaatat gaatctctga
     1501 aggacaaagc tcccgaggca attaattacg aacagatcaa gaaagatctg gctgaggaac
     1561 tgacattcga tatcgactat aagactagcg aggtgaacca gagggtcttt tccctggacg
     1621 aggtgtttga aatcgccaat ttcaacaatt acctgaacca gtccggcatt actaaattca
     1681 ataccatcat tggcgggaag tttgtgaacg gggagaatac caagcgcaag ggaattaacg
     1741 aatacatcaa tctgtatagc cagcagatca acgacaaaac tctgaagaaa tacaagatgt
     1801 ctgtgctgtt caaacagatc ctgagtgata ccgagtccaa gtcttttgtc attgataaac
     1861 tggaagatga ctcagacgtg gtcactacca tgcagagctt ttatgagcag atcgccgctt
     1921 tcaagacagt ggaggaaaaa tctattaagg aaactctgag tctgctgttc gatgacctga
     1981 aagcccagaa gctggacctg agtaagatct acttcaaaaa cgataagagt ctgacagacc
     2041 tgtcacagca ggtgtttgat gactattccg tgattgggac cgccgtcctg gagtacatta
     2101 cacagcagat cgctccaaag aacctggata atccctctaa gaaagagcag gaactgatcg
     2161 ctaagaaaac cgagaaggca aaatatctga gtctggaaac aattaagctg gcactggagg
     2221 agttcaacaa gcacagggat attgacaaac agtgccgctt tgaggaaatc ctggccaact
     2281 tcgcagccat ccccatgatt tttgatgaga tcgcccagaa caaagacaat ctggctcaga
     2341 tcagtattaa gtaccagaac cagggcaaga aagacctgct gcaggcttca gcagaagatg
     2401 acgtgaaagc catcaaggat ctgctggacc agaccaacaa tctgctgcac aagctgaaaa
     2461 tcttccatat tagtcagtca gaggataagg ctaatatcct ggataaagac gaacacttct
     2521 acctggtgtt cgaggaatgt tacttcgagc tggcaaacat tgtccccctg tataacaaga
     2581 ttaggaacta catcacacag aagccttact ctgacgagaa gtttaaactg aacttcgaaa
     2641 atagtaccct ggccaacggg tgggataaga acaaggagcc tgacaacaca gctatcctgt
     2701 tcatcaagga tgacaagtac tatctgggag tgatgaataa gaaaaacaat aagatcttcg
     2761 atgacaaagc cattaaggag aacaaagggg aaggatacaa gaaaatcgtg tataagctgc
     2821 tgcccggcgc aaataagatg ctgcctaagg tgttcttcag cgccaagagt atcaaattct
     2881 acaacccatc cgaggacatc ctgcggatta gaaatcactc aacacatact aagaacggga
     2941 gcccccagaa gggatatgag aaatttgagt tcaacatcga ggattgcagg aagtttattg
     3001 acttctacaa gcagagcatc tccaaacacc ctgaatggaa ggattttggc ttccggtttt
     3061 ccgacacaca gagatataac tctatcgacg agttctaccg cgaggtggaa aatcaggggt
     3121 ataagctgac ttttgagaac atttctgaaa gttacatcga cagcgtggtc aatcagggaa
     3181 agctgtacct gttccagatc tataacaaag atttttcagc atacagcaag ggcagaccaa
     3241 acctgcatac actgtactgg aaggccctgt tcgatgagag gaatctgcag gacgtggtct
     3301 ataaactgaa cggagaggcc gaactgtttt accggaagca gtctattcct aagaaaatca
     3361 ctcacccagc taaggaggcc atcgctaaca agaacaagga caatcctaag aaagagagcg
     3421 tgttcgaata cgatctgatt aaggacaagc ggttcaccga agataagttc tttttccatt
     3481 gtccaatcac cattaacttc aagtcaagcg gcgctaacaa gttcaacgac gagatcaatc
     3541 tgctgctgaa ggaaaaagca aacgatgtgc acatcctgag cattgaccga ggagagcggc
     3601 atctggccta ctataccctg gtggatggca aagggaatat cattaagcag gatacattca
     3661 acatcattgg caatgaccgg atgaaaacca actaccacga taaactggct gcaatcgaga
     3721 aggatagaga ctcagctagg aaggactgga agaaaatcaa caacattaag gagatgaagg
     3781 aaggctatct gagccaggtg gtccatgaga ttgcaaagct ggtcatcgaa tacaatgcca
     3841 ttgtggtgtt cgaggatctg aacttcggct ttaagagggg gcgctttaag gtggaaaaac
     3901 aggtctatca gaagctggag aaaatgctga tcgaaaagct gaattacctg gtgtttaaag
     3961 ataacgagtt cgacaagacc ggaggcgtcc tgagagccta ccagctgaca gctccctttg
     4021 aaactttcaa gaaaatggga aaacagacag gcatcatcta ctatgtgcca gccggattca
     4081 cttccaagat ctgccccgtg accggctttg tcaaccagct gtaccctaaa tatgagtcag
     4141 tgagcaagtc ccaggaattt ttcagcaagt tcgataagat ctgttataat ctggacaagg
     4201 ggtacttcga gttttccttc gattacaaga acttcggcga caaggccgct aaggggaaat
     4261 ggaccattgc ctccttcgga tctcgcctga tcaactttcg aaattccgat aaaaaccaca
     4321 attgggacac tagggaggtg tacccaacca aggagctgga aaagctgctg aaagactact
     4381 ctatcgagta tggacatggc gaatgcatca aggcagccat ctgtggcgag agtgataaga
     4441 aatttttcgc caagctgacc tcagtgctga atacaatcct gcagatgcgg aactcaaaga
     4501 ccgggacaga actggactat ctgattagcc ccgtggctga tgtcaacgga aacttcttcg
     4561 acagcagaca ggcacccaaa aatatgcctc aggatgcaga cgccaacggg gcctaccaca
     4621 tcgggctgaa gggactgatg ctgctgggcc ggatcaagaa caatcaggag gggaagaagc
     4681 tgaacctggt cattaagaac gaggaatact tcgagtttgt ccagaataga aataacggag
     4741 gtggaggttc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     4801 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4861 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4921 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4981 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     5041 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     5101 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     5161 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     5221 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     5281 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     5341 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     5401 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5461 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     5521 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5581 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5701 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     5761 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5821 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5881 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5941 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     6001 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     6061 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     6121 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     6181 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6241 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6301 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     6361 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6421 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6481 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6541 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6601 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6661 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     6721 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6781 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6841 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6901 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6961 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     7021 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     7081 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     7141 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7201 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     7261 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7321 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7381 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7441 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7501 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7561 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7681 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     7741 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     7801 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7861 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7921 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7981 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8041 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8101 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     8161 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8221 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     8281 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8341 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8401 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8461 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8521 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8581 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8641 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8701 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8761 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     8821 gcat
//
