LOCUS       Exported                6845 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-AsCas12f1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 12, 2021 from SnapGene 3.1.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6845
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        72..312
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    322..492
                     /note="sgRNA_v1"
     misc_feature    567..684
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        733..1243
                     /note="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1265..1864
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    2011..2132
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2551..3139)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(3310..4170)
                     /label=Ampicillin
                     /note="Ampicillin"
     CDS             complement(3310..4170)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4171..4275)
                     /gene="bla"
                     /note="AmpR promoter"
     primer_bind     4302..4318
                     /label=M13 rev
                     /note="M13 rev"
     polyA_signal    4389..4613
                     /label=bGH poly(A) signal
                     /note="bGH poly(A) signal"
     CDS             complement(4668..4688)
                     /label=SV40 NLS
                     /note="SV40 NLS"
     misc_feature    complement(4701..5966)
                     /note="AsCas12f human optimized"
     CDS             complement(5967..5987)
                     /label=SV40 NLS
                     /note="SV40 NLS"
     promoter        complement(6007..6025)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6067..6270)
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        6271..6650
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggtaccgatt agtgaacgga tctcgacggt atcgatcacg agactagcct cgagcggccg
       61 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      121 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      301 tgtggaaagg acgaaacacc gattcgtcgg ttcagcgacg ataagccgag aagtgccaat
      361 aaaactgtta agtggtttgg taacgctcgg taaggtagcc aaaaggctga aactccgtgc
      421 acaaagaccg cacggacgct tcacatatag ctcataaaca agggtttgcg agctagcttg
      481 tggagtgtga accgagaccc gagagagggt cggtctcctt tttttgaatt ctcgacctcg
      541 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
      601 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
      661 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
      721 ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg
      781 acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggactcgcac
      841 attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc
      901 ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg
      961 gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg
     1021 agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg
     1081 cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg
     1141 ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc
     1201 agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggagct
     1261 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     1321 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     1381 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     1441 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     1501 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     1561 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     1621 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     1681 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     1741 cctggaaacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     1801 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     1861 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     1921 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttctagagtc
     1981 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2041 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2101 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2161 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2221 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2281 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2341 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     2761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3421 atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc
     3481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcaggaaaca gctatgacca
     4321 tgattacgcc aagctctagc tagaggtcga cggtatcgag ccccagctgg ttctttccgc
     4381 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     4441 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     4501 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     4561 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
     4621 tgatcagcgg gtttaaactc aatggtgatg gtgatgatga ccggttagac tttcctcttc
     4681 ttcttgggag aaccaccaga cttaatgctc tcggcgatga tcttgtcgat gttggggatg
     4741 gcgatattcc gggcggcgtt gtagtcggcg ttggcctcgt agccgcaggc ccggcactta
     4801 aagatggcct ggccgattct gttgccggag tcgatgttgc cgcactcgga gcatctctgg
     4861 ctggtgtact gggggtcgat cttgatcact ttgatgccgg cctcctcggc cttgtagatg
     4921 atcttctgct gcaggtcgta gtaggtccag ttctgcagga atctgctgcc gatgtctctg
     4981 atgtttgtca gatcctccat ctggattgtg ccgcagccct ccttgatggc catgtccacg
     5041 atgtatctgc tgtaccggtg attggtggtg tctctgaaat tggcgatctt atccctcagc
     5101 tgctcaatgg gcttgattct cttgtctctg ccgtggccgc ccctggcgcc gccggcgtac
     5161 ttgccctgtc tcagcatgga gatgcgccgg ctctccacct gcctcctgaa gttctcaatc
     5221 tcgccgccct ccagcttgta cctggcgggg gtgtgctgga aggccatgta cacggccacg
     5281 gccacgccca ggtcaatgcc catgatcttg ttcaggtcca gcactctggt ctgtggctcg
     5341 aagtcgtagc tgatgttcag gtaccacttg ttcttccggt cgtcgtggat aatctggctg
     5401 gcgctcacct ggtactcgcc ggacaggatt ctgtccatga tggtcttgcc ggcgcccctc
     5461 acgatgatga tcacggagat ctttctcttc acgttcatct cctgcttggc ggggttgctc
     5521 agcagagaca ggctggcgat gtagtcgccg tgattcatcc tgttcacgga gatgttctcc
     5581 ttgatcaggt ccagggggat gtccctcttg tagctgggga tagacatgtc gccgcgcagg
     5641 atctccttct ggtaggcctt gaacctgtct gtggctctct tgatggtctg gctcagattt
     5701 ccagagttca gtctgtaggc ctttgtcttg atggtgtggt aggcgtagcc gtgcacgttg
     5761 gtgtagccca ggatgtcctt gctcttgggg tactcgccgt ggttatcctt gtagtcgctg
     5821 ctgaagccca tccactccca ggccagctgt gtggccttat tcagggcgaa tctggtttcc
     5881 tgctgcagct gtctcaggat ggtgccgaac tccttccagt ccaggtccag aggcttcacg
     5941 atctcgtatc tgtacacctt gatcatgact ttcctcttct tcttgggggc catggtggcg
     6001 gctctcccta tagtgagtcg tattagcggc cgcggatctc tagcggatct gacggttcac
     6061 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     6121 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     6181 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     6241 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     6301 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     6361 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     6421 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     6481 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     6541 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     6601 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata
     6661 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     6721 attgtcggtc aagaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     6781 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat
     6841 cgata
//
