LOCUS       Exported                7935 bp ds-DNA     circular SYN 06-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  AUTHORS   Dell
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 10, 2021 from SnapGene Viewer 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7935
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        683..960
                     /label=chicken beta-actin promoter
     CDS             1210..1230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1231..3597
                     /codon_start=1
                     /label=Cas13Y.1
                     /translation="NGIELKKEEAAFYFNQAELNLKAIEDNIFDKERRKTLLNNPQILA
                     KMENFIFNFRDVTKNAKGEIDCLLLKLRELRNFYSHYVHKRDVRELSKGEKPILEKYYQ
                     FAIESTGSENVKLEIIENDAWLADAGVLFFLCIFLKKSQANKLISGISGFKRNDDTGQP
                     RRNLFTYFSIREGYKVVPEMQKHFLLFSLVNHLSNQDDYIEKAHQPYDIGEGLFFHRIA
                     STFLNISGILRNMKFYTYQSKRLVEQRGELKREKDIFAWEEPFQGNSYFEINGHKGVIG
                     EDELKELCYAFLIGNQDANKVEGRITQFLEKFRNANSVQQVKDDEMLKPEYFPANYFAE
                     SGVGRIKDRVLNRLNKAIKSNKAKKGEIIAYDKMREVMAFINNSLPVDEKLKPKDYKRY
                     LGMVRFWDREKDNIKREFETKEWSKYLPSNFWTAKNLERVYGLAREKNAELFNKLKADV
                     EKMDERELEKYQKINDAKDLANLRRLASDFGVKWEEKDWDEYSGQIKKQITDSQKLTIM
                     KQRITAGLKKKHGIENLNLRITIDINKSRKAVLNRIAIPRGFVKRHILGWQESEKVSKK
                     IREAECEILLSKEYEELSKQFFQSKDYDKMTRINGLYEKNKLIALMAVYLMGQLRILFK
                     EHTKLDDITKTTVDFKISDKVTVKIPFSNYPSLVYTMSSKYVDNIGNYGFSNKDKDKPI
                     LGKIDVIEKQRMEFIKEVLGFEKYLFDDKIIDKSKFADTATHISFAEIVEELVEKGWDK
                     DRLTKLKDARNKALHGEILTGTSFDETKSLINELKK"
     CDS             3598..3618
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3625..3690
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3700..3834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3841..4081
                     /label=U6 Promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    4114..4149
                     /label=DR
     enhancer        4223..4526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4527..4730
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4765..5475
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    5492..5716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6138..6242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6243..7103
                     /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      7274..7862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tatcccctga ttctctcgag ttaaccctag aaagataatc atattgtgac gtacgttaaa
       61 gataatcatg cgtaaaattg acgcatgtgt tttatcgatc tgtatatcga ggtttattta
      121 ttaatttgaa tagatattaa gttttattat atttacactt acatactaat aataaattca
      181 acaaacaatt tatttatgtt tatttattta ttaaaaaaaa acaaaaactc aaaatttctt
      241 ctataaagta acaaaacttt taaacattct ctcttttaca aaaataaact tattttgtac
      301 tttaaaaaca gtcatgttgt attataaaat aagtaattag cttaacttat acataataga
      361 aacaaattat acttattaat taatctagag gtacccgtta cataacttac ggtaaatggc
      421 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
      481 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      541 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      601 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      661 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      721 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      781 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      841 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      901 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      961 ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
     1021 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     1081 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
     1141 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
     1201 gccaccatgc ccaagaagaa gcggaaggtg aatggcatcg agctgaagaa ggaagaagcc
     1261 gccttctact tcaatcaggc cgagctgaac ctgaaggcca ttgaggacaa catcttcgac
     1321 aaggagagac ggaaaacact gctgaacaac ccccagatcc tggccaagat ggagaacttt
     1381 atcttcaatt tccgggacgt gaccaagaac gccaagggcg aaatcgactg cctgctgctg
     1441 aagctgagag agctgcggaa cttttacagc cactacgtgc acaagcggga cgtcagagaa
     1501 ctgagcaagg gcgagaagcc gatcctggag aagtactacc agttcgccat cgaatccacc
     1561 ggctctgaga acgtgaagct cgaaatcatc gaaaacgacg cctggctggc cgacgccggc
     1621 gtgctgttct tcctgtgcat cttcctgaag aagagccagg caaacaagct gatcagcggc
     1681 atcagcggct tcaagagaaa cgacgacacc ggccagcctc ggagaaacct gttcacctac
     1741 ttctccatcc gggagggcta caaggtggtg cccgaaatgc agaagcactt cctgctgttc
     1801 tccctggtga accacctgag caaccaggac gattatatcg aaaaggccca ccagccctac
     1861 gacatcggcg agggcctctt cttccaccgg attgccagca ccttcctgaa catctccgga
     1921 atcctgagaa acatgaagtt ctacacctat cagagcaaga gactggtgga gcagagaggc
     1981 gagctgaagc gggaaaagga catcttcgcc tgggaagaac cgtttcaggg caattcctac
     2041 tttgagatca acggccacaa gggcgtgatt ggcgaggacg agctgaagga gctgtgctac
     2101 gccttcctga tcggcaacca ggacgccaac aaggtggagg gccggatcac ccagttcctg
     2161 gagaagttca gaaacgccaa cagcgtgcag caggtgaagg acgacgagat gctgaagcct
     2221 gaatatttcc ccgccaacta ctttgccgag agcggcgtgg gccggatcaa ggaccgggtg
     2281 ctgaacagac tgaacaaggc catcaagagc aacaaggcca agaagggcga gatcatcgcc
     2341 tatgacaaga tgagagaagt gatggctttc atcaataact ctctgcccgt ggacgagaag
     2401 ctgaagccca aggattacaa gagatacctg ggcatggtga gattctggga tagagaaaag
     2461 gacaatatca agcgcgagtt cgaaacgaag gagtggagca agtatctgcc ctccaacttc
     2521 tggaccgcca agaacctgga gagagtgtac ggactggccc gggaaaagaa cgcagagctg
     2581 tttaacaagc tgaaggccga cgtggagaag atggacgaaa gagagctgga aaagtatcag
     2641 aagatcaacg acgccaagga tctggccaac ctgcggcggc tggccagcga cttcggagtg
     2701 aagtgggagg agaaggattg ggacgagtac tccggccaga tcaagaagca gatcacagat
     2761 tcccagaagc tgaccatcat gaagcagaga atcacagccg gcctgaagaa gaagcacggc
     2821 atcgaaaacc tgaacctgag gatcaccatc gacatcaaca agtccagaaa ggccgtgctg
     2881 aatcggatcg ccatccccag aggatttgtg aagcggcaca tcctgggctg gcaggaatcc
     2941 gagaaggtga gcaagaagat cagagaagcc gaatgcgaga ttctgctgag caaggagtac
     3001 gaggagctga gcaagcagtt ctttcagagc aaggactacg acaagatgac ccgcatcaac
     3061 ggcctgtacg agaagaataa gctgatcgcc ctgatggccg tgtatctgat ggggcagctg
     3121 agaatcctgt tcaaggagca caccaagctg gacgacatca ccaagaccac cgtggatttc
     3181 aagatcagcg acaaggtgac cgtgaagatc cccttctcca actatccctc cctggtgtac
     3241 accatgagca gcaagtacgt ggacaatatc ggcaactacg gcttcagcaa caaggacaag
     3301 gataagccca ttctgggcaa gatcgacgtg atcgagaagc agcggatgga gtttatcaag
     3361 gaggtgctgg gattcgagaa gtacctgttt gacgataaga tcatcgacaa gagcaagttc
     3421 gccgacaccg ccacccacat cagctttgcc gaaatcgtgg aagaactggt ggagaagggc
     3481 tgggacaagg accggctgac gaagctgaag gatgcccgga acaaggccct gcacggcgag
     3541 atcctgaccg gcaccagctt cgacgagaca aagtccctga tcaacgagct gaagaagccc
     3601 aagaagaagc ggaaggtgtc ctctgattat aaggaccacg atggcgacta caaggatcac
     3661 gacatcgatt ataaggacga tgacgataag taaaagctta acttgtttat tgcagcttat
     3721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcactagt
     3841 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     3901 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     3961 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     4021 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     4081 cgaaacaccg atgtcttcct gggacgaaga caagctgtga tagacctcga tttgtggggt
     4141 agtaacagct ttttttacgc gttagttatt aatagtaatc aattacgggg tcattagttc
     4201 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4261 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4321 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4381 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4441 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4501 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4561 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4621 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4681 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4741 accgtcagat ccgctagcgc caccatggtg agcaagggcg aggaggataa catggccatc
     4801 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     4861 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     4921 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     4981 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     5041 cccgagggct tcaagtggga gcgcgtgatg aacttcgatg acggcggcgt ggtgaccgtg
     5101 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     5161 aacttcccct ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc
     5221 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     5281 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     5341 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     5401 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     5461 gagctgtaca agtaactctt ggcccggatc cctgtgcctt ctagttgcca gccatctgtt
     5521 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5581 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5641 ggggtggggc aggacagcaa gggggaggat tgggaaaaca atagcaggca tgctggggat
     5701 gcggtgggct ctatgggtcg acggcgcgcc ctagacgaag cagctccagc ctacacaatc
     5761 gctcaagacg tgtaatgctt ttattatata ttagtcacga tatctataac aagaaaatat
     5821 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     5881 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     5941 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     6001 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     6061 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta aggatccgca
     6121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6241 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6301 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6361 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6421 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6481 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6541 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6601 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6661 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6721 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6781 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6841 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6901 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6961 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7021 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7081 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7261 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7321 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7381 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7441 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7501 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7561 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7621 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7681 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7741 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7801 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7861 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7921 tgttctttcc tgcgt
//
