LOCUS       Exported                7975 bp ds-DNA     circular SYN 28-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pT3TS-spCas9-ABE-Umax-rest1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 29, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.2
FEATURES             Location/Qualifiers
     source          1..7975
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             772..864
                     /codon_start=1
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.6"
                     /note="/other=CDS"
                     /translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
     protein_bind    877..898
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        913..943
                     /label=lac promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    951..967
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     5'UTR           1055..1097
                     /label=Xenopus globin 5'-UTR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=5'UTR"
     CDS             1119..1175
                     /codon_start=1
                     /label=bp NLS
                     /translation="MKRTADGSEFESPKKKRKV"
     CDS             1182..1676
                     /codon_start=1
                     /label=tadA variant
                     /label=tadA
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=87.9"
                     /note="/match_length=98.8"
                     /note="/other=CDS"
                     /translation="MSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGW
                     NRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYSTFEPCVMCAGAMIHSRIGRVVF
                     GVRQSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRRVFNAQKKAQ
                     SS"
     misc_feature    1503..1505
                     /label=N>Q
     CDS             1677..1778
                     /codon_start=1
                     /label=X-ten linker
                     /translation="INSGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             1779..5876
                     /codon_start=1
                     /label=spCas9
                     /label=cas9
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=98.9"
                     /note="/match_length=99.9"
                     /note="/other=CDS"
                     /translation="DKKYSIGLAIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRLE
                     NLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGHK
                     PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY
                     YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPSE
                     EVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVA
                     QILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNA
                     VVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEI
                     TLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKES
                     ILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITI
                     MERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELA
                     LPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADAN
                     LDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDA
                     TLIHQSITGLYETRIDLSQLGGD"
     CDS             5877..5930
                     /codon_start=1
                     /label=bp NLS
                     /translation="KRTADGSEFESPKKKRKV"
     3'UTR           5955..6080
                     /label=Xenopus globin 3'-UTR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=3'UTR"
     promoter        complement(6175..6193)
                     /label=T7 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             6196..6353
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=90.8"
                     /note="/other=CDS"
                     /translation="IHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPN
                     SCAA*MA"
     rep_origin      complement(6385..6813)
                     /direction=LEFT
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     promoter        6840..6944
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             6945..7805
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.7"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPLFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag aagctcagaa
     1081 taaacgctca actttggcag atctaactcg acgccaccat gaaaagaact gcagatggat
     1141 ccgagttcga gagccccaag aaaaagagaa aggtgggaag catgagcgaa gtcgagtttt
     1201 cacacgaata ctggatgaga catgctctca cactcgcaaa gagggctaga gatgagaggg
     1261 aggttcctgt gggcgcagtg ctggtcctga ataacagggt tataggcgag ggatggaacc
     1321 gcgcaattgg actgcatgac ccaaccgcac acgcagagat tatggcactc aggcaaggag
     1381 gactggtcat gcagaactat agactcattg acgccacact gtatagcacc ttcgagcctt
     1441 gtgtgatgtg cgcaggagcc atgatccaca gtaggatcgg tagagtcgtg tttggagtta
     1501 gacagtcaaa gcgcggagct gccggcagtc tgatgaatgt tctgaactac ccaggaatga
     1561 atcatagggt cgagatcacc gaaggcattc tggccgacga atgtgccgct ctgctgtgcg
     1621 atttctacag aatgccaaga cgggttttca acgctcagaa gaaagcccag tcttccatta
     1681 atagcggagg ttccagcgga ggctccagtg gaagcgaaac acctgggaca tcagagtctg
     1741 ccactccaga gtccagtgga ggctcctctg gcggcagtga taagaagtat agcatcggcc
     1801 tggctattgg aactaactcc gtgggttggg cagtgattac agacgactac aaggtcccta
     1861 gcaagaaatt taaggtgctg ggtaacaccg acaggcacag catcaagaaa aatctgattg
     1921 gagccctgct gttcggttct ggagagactg ccgaagcaac acgcctgaaa agaacagcaa
     1981 gaaggcgcta taccagaagg aagaatagaa tctgttacct gcaggagatt ttctctaacg
     2041 aaatggctaa ggtggacgat tcattctttc ataggctgga ggaaagtttc ctggtcgagg
     2101 aagataagaa acacgagcgc catcctatct ttggaaacat tgtggacgag gtcgcctatc
     2161 acgaaaaata cccaaccatc tatcatctgc gcaagaaact ggctgactct actgataaag
     2221 ccgacctgag actgatctat ctggctctgg cccacatgat taagttcagg ggtcattttc
     2281 tgatcgaggg cgatctgaac cccgacaatt ccgatgtgga caagctgttc atccagctgg
     2341 tccagattta caatcagctg tttgaggaaa accctattaa tgcttccaga gtggacgcaa
     2401 aagctatcct gtcagccagg ctgtccaagt cacgcagact ggagaacctg attgcacagc
     2461 tgcccggaga aaagaggaac ggtctgtttg gaaatctgat cgctctgagt ctgggcctga
     2521 ctcctaactt caaaagcaat tttgatctgg ctgaggacgc caaactgcag ctgtcaaagg
     2581 acacatatga cgatgacctg gataacctgc tggcacagat cggagatcag tacgctgacc
     2641 tgttcctggc tgccaaaaat ctgtccgacg caatcctgct gtcagatatt ctgagagtga
     2701 acagcgagat tacaaaagct cctctgagtg ccagcatgat caagagatat gacgagcacc
     2761 atcaggatct gaccctgctg aaggctctgg tcaggcagca gctgccagag aagtacaagg
     2821 aaattttctt tgatcagtcc aagaacggct acgccggtta tatcgacgga ggcgcatcac
     2881 aggaggaatt ctacaagttt atcaaaccta ttctggagaa gatggacgga actgaggaac
     2941 tgctggtgaa actgaataga gaggacctgc tgaggaagca gcgcacattt gataacggtt
     3001 ccatcccaca ccagattcat ctgggagagc tgcacgctat cctgaggcgc caggaagact
     3061 tctacccctt tctgaaagat aaccgcgaga agatcgaaaa aattctgacc ttcagaatcc
     3121 cttactatgt gggtccactg gctcgcggaa acagcagatt tgcctggatg actcgcaaat
     3181 ccgaggaaac cattactcct tggaacttcg aggaagtggt cgataagggc gcctctgcac
     3241 agtccttcat cgagagaatg actaattttg acaaaaacct gcccaatgag aaagtgctgc
     3301 ctaagcactc cctgctgtac gagtatttca ctgtctataa cgaactgaca aaggtgaaat
     3361 acgtcaccga gggcatgaga aagccagcct tcctgtcagg agagcagaag aaagcaatcg
     3421 tggatctgct gtttaaaacc aacaggaaag tgactgtcaa gcagctgaag gaggactact
     3481 tcaagaaaat tgaatgcttc gattccgtgg agatcagcgg agtcgaagac agatttaacg
     3541 caagcctggg cgcttaccac gatctgctga agatcattaa ggataaagac ttcctggaca
     3601 acgaggaaaa tgaggatatc ctggaagaca ttgtgctgac actgaccctg tttgaggaca
     3661 gaggaatgat cgaggaaaga ctgaaaacct atgctcatct gttcgatgac aaggtgatga
     3721 aacagctgaa gagaaggcgc tacactggct ggggtagact gagcaggaag ctgatcaacg
     3781 gcattaggga taaacagtca ggaaagacaa tcctggactt tctgaaaagt gatggcttcg
     3841 ccaaccgcaa ttttatgcag ctgattcacg atgacagtct gaccttcaaa gaggacatcc
     3901 agaaggctca ggtgtctgga cagggccact ccctgcatga gcagattgca aacctggctg
     3961 gaagcccagc catcaagaaa ggcattctgc agacagtgaa aatcgtcgat gagctggtga
     4021 aagtcatggg ccataagccc gaaaacatcg tgattgagat ggctcgcgaa aatcagacaa
     4081 cccagaaggg tcagaagaac agtagagaga ggatgaaaag aatcgaggaa ggcattaagg
     4141 agctgggtag ccagatcctg aaagagcacc cagtggaaaa cacacagctg cagaatgaga
     4201 agctgtatct gtactatctg cagaatggaa gagatatgta cgtggaccag gagctggata
     4261 ttaacaggct gtctgattac gacgtggatc atatcgtccc ccagagtttc atcaaagatg
     4321 acagcattga caacaaggtg ctgaccaggt ccgacaaaaa cagaggaaaa tcagataatg
     4381 tccctagtga ggaagtggtc aagaaaatga agaactactg gagacagctg ctgaatgcca
     4441 aactgatcac tcagaggaag tttgataacc tgacaaaagc agagcgcggt ggactgtcag
     4501 aactggacaa agctggattc atcaagaggc agctggtgga aacacgccag atcactaaac
     4561 acgtcgcaca gattctggat agtcgcatga acacaaagta cgatgagaat gacaaactga
     4621 tcagagaagt gaaggtcatt accctgaaga gtaaactggt cagcgacttt aggaaagatt
     4681 tccagtttta taaggtccgc gagattaaca attatcacca tgcccatgac gcatacctga
     4741 acgccgtggt cggtaccgca ctgatcaaga aatacccaaa actggagagc gaattcgtgt
     4801 acggagacta taaggtgtac gatgtcagaa aaatgatcgc caagtccgag caggaaattg
     4861 gaaaagctac tgccaagtat ttcttttact caaacatcat gaatttcttt aagacagaga
     4921 tcaccctggc caatggagaa atccgcaaaa ggcccctgat tgagacaaac ggagagacag
     4981 gcgaaatcgt gtgggacaaa ggcagagatt ttgcaaccgt gaggaaggtc ctgagcatgc
     5041 ctcaagtgaa tatcgtcaag aaaactgagg tgcagacagg cggtttctca aaagaaagta
     5101 ttctgccaaa acgcaactct gataagctga tcgctagaaa gaaagactgg gaccctaaga
     5161 agtatggagg ctttgactct cccactgtgg catactccgt cctggtggtc gctaaggtgg
     5221 agaagggcaa aagcaagaaa ctgaaatctg tcaaggagct gctgggtatc acaattatgg
     5281 agagaagctc tttcgagaag aacccaatcg attttctgga ggccaaaggt tataaggaag
     5341 tgaagaaaga cctgatcatt aaactgccca agtacagtct gtttgagctg gaaaacggca
     5401 ggaaacgcat gctggcaagc gctggagagc tgcagaaagg caatgaactg gccctgcctt
     5461 ctaagtacgt gaacttcctg tatctggcaa gccactacga gaagctgaaa ggatctccag
     5521 aggataacga acagaaacag ctgtttgtgg agcagcacaa gcattatctg gacgagatca
     5581 ttgaacagat tagcgagttc tctaaaagag tgatcctggc cgacgcaaat ctggataagg
     5641 tcctgtctgc ttacaacaaa cacagagata agcccatcag ggagcaggcc gaaaatatca
     5701 ttcatctgtt cactctgaca aacctgggcg cacctgcagc tttcaagtac ttcgacacta
     5761 caatcgatag aaagaggtac acctccacta aggaggtgct ggacgctaca ctgatccatc
     5821 agagtattac cggcctgtac gaaacaagga ttgacctgtc tcagctgggt ggcgacaaaa
     5881 gaactgcaga tggatccgag ttcgagagcc ccaagaaaaa gagaaaggtg tgaccgcgga
     5941 tctggttacc actaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata
     6001 ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa
     6061 taaaaagaaa gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6121 aacccccccc cccccctgca gggccaagtc ggcctctaga ggtacccaat tcgccctata
     6181 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     6241 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     6301 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcaaa
     6361 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6421 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6481 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6541 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6601 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6661 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6721 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6781 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     6841 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatctg ctcatgagac
     6901 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6961 tccgtgtcgc ccttattccc ctttttgcgg cattttgcct tcctgttttt gctcacccag
     7021 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7081 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7141 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7201 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7261 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7321 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7381 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7441 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7501 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7561 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7621 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7681 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7741 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7801 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7861 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7921 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
