LOCUS       Exported                7517 bp ds-DNA     circular SYN 28-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   <unknown
VERSION     .
KEYWORDS    pT3TS-spCas9 ABE-Umax-ex2-rest1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7517)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 29, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7517
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /label=ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=99.83"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             771..864
                     /codon_start=1
                     /label=lacI
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.679593721144968"
                     /note="/other=CDS"
                     /translation="SAQYANRLSPRVGRFINAAGTTGFPTGKRAV"
     protein_bind    complement(877..898)
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=addgene"
                     /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=addgene"
                     /note="/fragment=False"
                     /note="/identity=96.875"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    complement(947..969)
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=95.83333333333334"
                     /note="/other=protein bind"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /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=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=5'UTR"
     CDS             1119..1181
                     /codon_start=1
                     /label=bpNLS
                     /translation="MKRTADGSEFESPKKKRKVGS"
     CDS             1188..3560
                     /codon_start=1
                     /label=△HNHN-cas9
                     /translation="DKKYSIGLAIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRLE
                     NLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGHK
                     PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGS"
     misc_feature    3570..4067
                     /label=tadA variant
     misc_feature    3888..3890
                     /label=N>Q
     misc_feature    4077..5426
                     /label=Delta-HNH-Cas9
     misc_feature    5427..5480
                     /label=bpNLS
     3'UTR           5505..5630
                     /label=Xenopus globin 3'-UTR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=98.438"
                     /note="/match_length=100.0"
                     /note="/other=3'UTR"
     promoter        complement(5717..5735)
                     /label=T7 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             5738..5899
                     /codon_start=1
                     /label=lacZ
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=5.2682926829268295"
                     /note="/other=CDS"
                     /translation="IHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPN
                     SCAA*MANG"
     primer_bind     complement(5742..5758)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5900..6355)
                     /direction=LEFT
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     promoter        6382..6486
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             6487..7347
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=99.884"
                     /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 gccaactctt 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 catgagcgat aagaagtata
     1201 gcatcggcct ggctattgga actaactccg tgggttgggc agtgattaca gacgactaca
     1261 aggtccctag caagaaattt aaggtgctgg gtaacaccga caggcacagc atcaagaaaa
     1321 atctgattgg agccctgctg ttcggttctg gagagactgc cgaagcaaca cgcctgaaaa
     1381 gaacagcaag aaggcgctat accagaagga agaatagaat ctgttacctg caggagattt
     1441 tctctaacga aatggctaag gtggacgatt cattctttca taggctggag gaaagtttcc
     1501 tggtcgagga agataagaaa cacgagcgcc atcctatctt tggaaacatt gtggacgagg
     1561 tcgcctatca cgaaaaatac ccaaccatct atcatctgcg caagaaactg gctgactcta
     1621 ctgataaagc cgacctgaga ctgatctatc tggctctggc ccacatgatt aagttcaggg
     1681 gtcattttct gatcgagggc gatctgaacc ccgacaattc cgatgtggac aagctgttca
     1741 tccagctggt ccagatttac aatcagctgt ttgaggaaaa ccctattaat gcttccagag
     1801 tggacgcaaa agctatcctg tcagccaggc tgtccaagtc acgcagactg gagaacctga
     1861 ttgcacagct gcccggagaa aagaggaacg gtctgtttgg aaatctgatc gctctgagtc
     1921 tgggcctgac tcctaacttc aaaagcaatt ttgatctggc tgaggacgcc aaactgcagc
     1981 tgtcaaagga cacatatgac gatgacctgg ataacctgct ggcacagatc ggagatcagt
     2041 acgctgacct gttcctggct gccaaaaatc tgtccgacgc aatcctgctg tcagatattc
     2101 tgagagtgaa cagcgagatt acaaaagctc ctctgagtgc cagcatgatc aagagatatg
     2161 acgagcacca tcaggatctg accctgctga aggctctggt caggcagcag ctgccagaga
     2221 agtacaagga aattttcttt gatcagtcca agaacggcta cgccggttat atcgacggag
     2281 gcgcatcaca ggaggaattc tacaagttta tcaaacctat tctggagaag atggacggaa
     2341 ctgaggaact gctggtgaaa ctgaatagag aggacctgct gaggaagcag cgcacatttg
     2401 ataacggttc catcccacac cagattcatc tgggagagct gcacgctatc ctgaggcgcc
     2461 aggaagactt ctaccccttt ctgaaagata accgcgagaa gatcgaaaaa attctgacct
     2521 tcagaatccc ttactatgtg ggtccactgg ctcgcggaaa cagcagattt gcctggatga
     2581 ctcgcaaatc cgaggaaacc attactcctt ggaacttcga ggaagtggtc gataagggcg
     2641 cctctgcaca gtccttcatc gagagaatga ctaattttga caaaaacctg cccaatgaga
     2701 aagtgctgcc taagcactcc ctgctgtacg agtatttcac tgtctataac gaactgacaa
     2761 aggtgaaata cgtcaccgag ggcatgagaa agccagcctt cctgtcagga gagcagaaga
     2821 aagcaatcgt ggatctgctg tttaaaacca acaggaaagt gactgtcaag cagctgaagg
     2881 aggactactt caagaaaatt gaatgcttcg attccgtgga gatcagcgga gtcgaagaca
     2941 gatttaacgc aagcctgggc gcttaccacg atctgctgaa gatcattaag gataaagact
     3001 tcctggacaa cgaggaaaat gaggatatcc tggaagacat tgtgctgaca ctgaccctgt
     3061 ttgaggacag aggaatgatc gaggaaagac tgaaaaccta tgctcatctg ttcgatgaca
     3121 aggtgatgaa acagctgaag agaaggcgct acactggctg gggtagactg agcaggaagc
     3181 tgatcaacgg cattagggat aaacagtcag gaaagacaat cctggacttt ctgaaaagtg
     3241 atggcttcgc caaccgcaat tttatgcagc tgattcacga tgacagtctg accttcaaag
     3301 aggacatcca gaaggctcag gtgtctggac agggccactc cctgcatgag cagattgcaa
     3361 acctggctgg aagcccagcc atcaagaaag gcattctgca gacagtgaaa atcgtcgatg
     3421 agctggtgaa agtcatgggc cataagcccg aaaacatcgt gattgagatg gctcgcgaaa
     3481 atcagacaac ccagaagggt cagaagaaca gtagagagag gatgaaaaga atcgaggaag
     3541 gcattaagga gctgggtagc ggaggtagta gcgaagtcga gttttcacac gaatactgga
     3601 tgagacatgc tctcacactc gcaaagaggg ctagagatga gagggaggtt cctgtgggcg
     3661 cagtgctggt cctgaataac agggttatag gcgagggatg gaaccgcgca attggactgc
     3721 atgacccaac cgcacacgca gagattatgg cactcaggca aggaggactg gtcatgcaga
     3781 actatagact cattgacgcc acactgtata gcaccttcga gccttgtgtg atgtgcgcag
     3841 gagccatgat ccacagtagg atcggtagag tcgtgtttgg agttagacag tcaaagcgcg
     3901 gagctgccgg cagtctgatg aatgttctga actacccagg aatgaatcat agggtcgaga
     3961 tcaccgaagg cattctggcc gacgaatgtg ccgctctgct gtgcgatttc tacagaatgc
     4021 caagacgggt tttcaacgct cagaagaaag cccagtcttc cattaattcc ggaggcaggc
     4081 agctggtgga aacacgccag atcactaaac acgtcgcaca gattctggat agtcgcatga
     4141 acacaaagta cgatgagaat gacaaactga tcagagaagt gaaggtcatt accctgaaga
     4201 gtaaactggt cagcgacttt aggaaagatt tccagtttta taaggtccgc gagattaaca
     4261 attatcacca tgcccatgac gcatacctga acgccgtggt cggtaccgca ctgatcaaga
     4321 aatacccaaa actggagagc gaattcgtgt acggagacta taaggtgtac gatgtcagaa
     4381 aaatgatcgc caagtccgag caggaaattg gaaaagctac tgccaagtat ttcttttact
     4441 caaacatcat gaatttcttt aagacagaga tcaccctggc caatggagaa atccgcaaaa
     4501 ggcccctgat tgagacaaac ggagagacag gcgaaatcgt gtgggacaaa ggcagagatt
     4561 ttgcaaccgt gaggaaggtc ctgagcatgc ctcaagtgaa tatcgtcaag aaaactgagg
     4621 tgcagacagg cggtttctca aaagaaagta ttctgccaaa acgcaactct gataagctga
     4681 tcgctagaaa gaaagactgg gaccctaaga agtatggagg ctttgactct cccactgtgg
     4741 catactccgt cctggtggtc gctaaggtgg agaagggcaa aagcaagaaa ctgaaatctg
     4801 tcaaggagct gctgggtatc acaattatgg agagaagctc tttcgagaag aacccaatcg
     4861 attttctgga ggccaaaggt tataaggaag tgaagaaaga cctgatcatt aaactgccca
     4921 agtacagtct gtttgagctg gaaaacggca ggaaacgcat gctggcaagc gctggagagc
     4981 tgcagaaagg caatgaactg gccctgcctt ctaagtacgt gaacttcctg tatctggcaa
     5041 gccactacga gaagctgaaa ggatctccag aggataacga acagaaacag ctgtttgtgg
     5101 agcagcacaa gcattatctg gacgagatca ttgaacagat tagcgagttc tctaaaagag
     5161 tgatcctggc cgacgcaaat ctggataagg tcctgtctgc ttacaacaaa cacagagata
     5221 agcccatcag ggagcaggcc gaaaatatca ttcatctgtt cactctgaca aacctgggcg
     5281 cacctgcagc tttcaagtac ttcgacacta caatcgatag aaagaggtac acctccacta
     5341 aggaggtgct ggacgctaca ctgatccatc agagtattac cggcctgtac gaaacaagga
     5401 ttgacctgtc tcagctgggt ggcgacaaaa gaactgcaga tggatccgag ttcgagagcc
     5461 ccaagaaaaa gagaaaggtg tgaccgcgga tctggttacc actaaaccag cctcaagaac
     5521 acccgaatgg agtctctaag ctacataata ccaacttaca ctttacaaaa tgttgtcccc
     5581 caaaatgtag ccattcgtat ctgctcctaa taaaaagaaa gtttcttcac attctaaaaa
     5641 aaaaaaaaaa aaaaaaaaaa aaaaaacccc cccccccctg cagggccaag tcggcctcta
     5701 gaggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     5761 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5821 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5881 agcctgaatg gcgaatggca aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5941 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6001 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6061 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6121 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6181 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6241 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6301 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     6361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6421 aatatgtatc tgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6481 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccctttttgc ggcattttgc
     6541 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6601 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6661 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6721 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6781 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6841 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6901 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6961 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7021 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7081 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7141 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7201 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7261 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7501 aagatcaaag gatcttc
//
