LOCUS       Exported                6954 bp DNA     circular SYN 07-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6954)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 27, 2022 from SnapGene Viewer 6.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..22
                     /label=S-BPK-1F
     promoter        27..43
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    46..70
                     /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)."
     CDS             114..137
                     /codon_start=1
                     /label=NLS
                     /translation="MPKKKRKV"
     CDS             144..3305
                     /codon_start=1
                     /label=Cas12i2
                     /translation="MSSAIKSYKSVLRPNERKNQLLKSTIQCLEDGSAFFFKMLQGLFG
                     GITPEIVRFSTEQEKQQQDIALWCAVNWFRPVSQDSLTHTIASDNLVEKFEEYYGGTAS
                     DAIKQYFSASIGESYYWNDCRQQYYDLCRELGVEVSDLTHDLEILCREKCLAVATESNQ
                     NNSIISVLFGTGEKEDRSVKLRITKKILEAISNLKEIPKNVAPIQEIILNVAKATKETF
                     RQVYAGNLGAPSTLEKFIAKDGQKEFDLKKLQTDLKKVIRGKSKERDWCCQEELRSYVE
                     QNTIQYDLWAWGEMFNKAHTALKIKSTRNYNFAKQRLEQFKEIQSLNNLLVVKKLNDFF
                     DSEFFSGEETYTICVHHLGGKDLSKLYKAWEDDPADPENAIVVLCDDLKNNFKKEPIRN
                     ILRYIFTIRQECSAQDILAAAKYNQQLDRYKSQKANPSVLGNQGFTWTNAVILPEKAQR
                     NDRPNSLDLRIWLYLKLRHPDGRWKKHHIPFYDTRFFQEIYAAGNSPVDTCQFRTPRFG
                     YHLPKLTDQTAIRVNKKHVKAAKTEARIRLAIQQGTLPVSNLKITEISATINSKGQVRI
                     PVKFDVGRQKGTLQIGDRFCGYDQNQTASHAYSLWEVVKEGQYHKELGCFVRFISSGDI
                     VSITENRGNQFDQLSYEGLAYPQYADWRKKASKFVSLWQITKKNKKKEIVTVEAKEKFD
                     AICKYQPRLYKFNKEYAYLLRDIVRGKSLVELQQIRQEIFRFIEQDCGVTRLGSLSLST
                     LETVKAVKGIIYSYFSTALNASKNNPISDEQRKEFDPELFALLEKLELIRTRKKKQKVE
                     RIANSLIQTCLENNIKFIRGEGDLSTTNNATKKKANSRSMDWLARGVFNKIRQLAPMHN
                     ITLFGCGSLYTSHQDPLVHRNPDKAMKCRWAAIPVKDIGDWVLRKLSQNLRAKNIGTGE
                     YYHQGVKEFLSHYELQDLEEELLKWRSDRKSNIPCWVLQNRLAEKLGNKEAVVYIPVRG
                     GRIYFATHKVATGAVSIVFDQKQVWVCNADHVAAANIALTVKGIGEQSSDEENPDGSRI
                     KLQLTS"
     CDS             3312..3341
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        3410..3426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3450..3497
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3671..3690)
                     /label=S-BPK-1R
     CDS             complement(3718..4377)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4378..4480)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene"
     rep_origin      complement(5006..5550)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the colE1 origin.
                     Correction: Low-copy-number p15A origin of replication 
                     (10~12)
                     "
     CDS             complement(5775..6857)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6858..6935)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat catatgccca
      121 agaagaagag gaaggttccc gggatgagca gcgccatcaa gagctacaag agcgtgctgc
      181 gccccaacga gcgcaagaac cagctgctga agagcaccat ccagtgcctg gaggacggca
      241 gcgccttctt cttcaagatg ctgcagggcc tgttcggcgg catcaccccc gagatcgtgc
      301 gcttcagcac cgagcaggag aagcagcagc aggacatcgc cctgtggtgc gccgtgaact
      361 ggttccgccc cgtgagccag gacagcctga cccacaccat cgccagcgac aacctggtgg
      421 agaagttcga ggagtactac ggcggcaccg ccagcgacgc catcaagcag tacttcagcg
      481 ccagcatcgg cgagagctac tactggaacg actgccgcca gcagtactac gacctgtgcc
      541 gcgagctggg cgtggaggtg agcgacctga cccacgacct ggagatcctg tgccgcgaga
      601 agtgcctggc cgtggccacc gagagcaacc agaacaacag catcatcagc gtgctgttcg
      661 gcaccggcga gaaggaggac cgcagcgtga agctgcgcat caccaagaag atcctggagg
      721 ccatcagcaa cctgaaggag atccccaaga acgtggcccc catccaggag atcatcctga
      781 acgtggccaa ggccaccaag gagaccttcc gccaggtgta cgccggcaac ctgggcgccc
      841 ccagcaccct ggagaagttc atcgccaagg acggccagaa ggagttcgac ctgaagaagc
      901 tgcagaccga cctgaagaag gtgatccgcg gcaagagcaa ggagcgcgac tggtgctgcc
      961 aggaggagct gcgcagctac gtggagcaga acaccatcca gtacgacctg tgggcctggg
     1021 gcgagatgtt caacaaggcc cacaccgccc tgaagatcaa gagcacccgc aactacaact
     1081 tcgccaagca gcgcctggag cagttcaagg agatccagag cctgaacaac ctgctggtgg
     1141 tgaagaagct gaacgacttc ttcgacagcg agttcttcag cggcgaggag acctacacca
     1201 tctgcgtgca ccacctgggc ggcaaggacc tgagcaagct gtacaaggcc tgggaggacg
     1261 accccgccga ccccgagaac gccatcgtgg tgctgtgcga cgacctgaag aacaacttca
     1321 agaaggagcc catccgcaac atcctgcgct acatcttcac catccgccag gagtgcagcg
     1381 cccaggacat cctggccgcc gccaagtaca accagcagct ggaccgctac aagagccaga
     1441 aggccaaccc cagcgtgctg ggcaaccagg gcttcacctg gaccaacgcc gtgatcctgc
     1501 ccgagaaggc ccagcgcaac gaccgcccca acagcctgga cctgcgcatc tggctgtacc
     1561 tgaagctgcg ccaccccgac ggccgctgga agaagcacca catccccttc tacgacaccc
     1621 gcttcttcca ggagatctac gccgccggca acagccccgt ggacacctgc cagttccgca
     1681 ccccccgctt cggctaccac ctgcccaagc tgaccgacca gaccgccatc cgcgtgaaca
     1741 agaagcacgt gaaggccgcc aagaccgagg cccgcatccg cctggccatc cagcagggca
     1801 ccctgcccgt gagcaacctg aagatcaccg agatcagcgc caccatcaac agcaagggcc
     1861 aggtgcgcat ccccgtgaag ttcgacgtgg gccgccagaa gggcaccctg cagatcggcg
     1921 accgcttctg cggctacgac cagaaccaga ccgccagcca cgcctacagc ctgtgggagg
     1981 tggtgaagga gggccagtac cacaaggagc tgggctgctt cgtgcgcttc atcagcagcg
     2041 gcgacatcgt gagcatcacc gagaaccgcg gcaaccagtt cgaccagctg agctacgagg
     2101 gcctggccta cccccagtac gccgactggc gcaagaaggc cagcaagttc gtgagcctgt
     2161 ggcagatcac caagaagaac aagaagaagg agatcgtgac cgtggaggcc aaggagaagt
     2221 tcgacgccat ctgcaagtac cagccccgcc tgtacaagtt caacaaggag tacgcctacc
     2281 tgctgcgcga catcgtgcgc ggcaagagcc tggtggagct gcagcagatc cgccaggaga
     2341 tcttccgctt catcgagcag gactgcggcg tgacccgcct gggcagcctg agcctgagca
     2401 ccctggagac cgtgaaggcc gtgaagggca tcatctacag ctacttcagc accgccctga
     2461 acgccagcaa gaacaacccc atcagcgacg agcagcgcaa ggagttcgac cccgagctgt
     2521 tcgccctgct ggagaagctg gagctgatcc gcacccgcaa gaagaagcag aaggtggagc
     2581 gcatcgccaa cagcctgatc cagacctgcc tggagaacaa catcaagttc atccgcggcg
     2641 agggcgacct gagcaccacc aacaacgcca ccaagaagaa ggccaacagc cgcagcatgg
     2701 actggctggc ccgcggcgtg ttcaacaaga tccgccagct ggcccccatg cacaacatca
     2761 ccctgttcgg ctgcggcagc ctgtacacca gccaccagga ccccctggtg caccgcaacc
     2821 ccgacaaggc catgaagtgc cgctgggccg ccatccccgt gaaggacatc ggcgactggg
     2881 tgctgcgcaa gctgagccag aacctgcgcg ccaagaacat cggcaccggc gagtactacc
     2941 accagggcgt gaaggagttc ctgagccact acgagctgca ggacctggag gaggagctgc
     3001 tgaagtggcg cagcgaccgc aagagcaaca tcccctgctg ggtgctgcag aaccgcctgg
     3061 ccgagaagct gggcaacaag gaggccgtgg tgtacatccc cgtgcgcggc ggccgcatct
     3121 acttcgccac ccacaaggtg gccaccggcg ccgtgagcat cgtgttcgac cagaagcagg
     3181 tgtgggtgtg caacgccgac cacgtggccg ccgccaacat cgccctgacc gtgaagggca
     3241 tcggcgagca gagcagcgac gaggagaacc ccgacggcag ccgcatcaag ctgcagctga
     3301 ccagcgctag tcatcaccat caccaccatc atcaccatca ctaggcggcc gcataatgct
     3361 taagtcgaac agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc
     3421 actataggga attcggtacc tgagaataac tagcataacc ccttggggcc tctaaacggg
     3481 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     3541 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     3601 ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc
     3661 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta
     3721 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     3781 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     3841 ttgcgtataa tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac
     3901 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     3961 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     4021 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     4081 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     4141 gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat
     4201 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     4261 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     4321 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     4381 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     4441 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     4501 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     4561 atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact
     4621 gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc
     4681 cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc
     4741 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     4801 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     4861 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     4921 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     4981 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     5041 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     5101 ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     5161 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     5221 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     5281 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     5341 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     5401 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     5461 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     5521 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     5581 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     5641 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     5701 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     5761 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5821 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     5881 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     5941 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     6001 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     6061 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     6121 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     6181 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     6241 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     6301 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     6361 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     6421 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     6481 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     6541 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     6601 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     6661 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     6721 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     6781 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     6841 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     6901 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcc
//
