LOCUS       MZB222        6209 bp DNA     circular SYN 14-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6209)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    208..226
                     /label=T7
                     /note="/vntifkey=21"
     protein_bind    227..251
                     /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)."
     RBS             265..287
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             301..330
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    337..630
                     /label=MZB-SUMO
     primer_bind     537..559
                     /label=MZB-SUMO 5'F
     primer_bind     complement(541..563)
                     /label=SUMO 5' Rev
     primer_bind     complement(609..651)
                     /label=oMB497
     primer_bind     complement(609..630)
                     /label=SUMO HiFi Rev
     primer_bind     636..693
                     /label=oMB499
     primer_bind     677..726
                     /label=oMB498
     misc_feature    697..1092
                     /label=Original
     primer_bind     complement(1071..1114)
                     /label=oMB500
     primer_bind     1099..1153
                     /label=oMB502
     primer_bind     complement(1140..1187)
                     /label=oMB503
     primer_bind     1171..1221
                     /label=oMB504
     primer_bind     1201..1256
                     /label=oMB501
     primer_bind     complement(1206..1239)
                     /label=oMB505
     primer_bind     1228..1256
                     /label=SUMO HiFi For
     primer_bind     complement(1247..1276)
                     /label=oMB404
     CDS             1320..1337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1389..1407)
                     /label=T7-Term
     terminator      1403..1450
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1486..1938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    complement(2029..2838)
                     /label=KanR2
                     /note="/vntifkey=21"
     misc_feature    complement(2943..3557)
                     /label=PBR322 Origin
                     /label=PBR322_Origin
                     /note="/vntifkey=21"
     rep_origin      2959..3542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3727..3865
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(3966..4156)
                     /label=ROP
                     /note="/vntifkey=21"
     misc_feature    complement(4627..4900)
                     /label=tet
                     /note="/vntifkey=21"
     misc_feature    complement(4958..6041)
                     /label=lacI
                     /note="/vntifkey=21"
     promoter        complement(6033..6109)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      181 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      241 ataacaattc cccttagaaa taattttgtt taactttaag aaggagatat acatatgggc
      301 catcaccatc accatcacca tcaccatcac ggcagcatga gcgatagcga agtgaaccag
      361 gaagcgaaac cggaagtgaa accggaagtg aaaccggaaa cccatattaa cctgaaagtg
      421 agcgatggca gcagcgaaat tttttttaaa attaaaaaaa ccaccccgct gcgtcgtctg
      481 atggaagcgt ttgcgaaacg tcagggcaaa gaaatggata gcctgcgttt tctgtatgat
      541 ggcattcgta ttcaggcgga tcagaccccg gaagatctgg atatggaaga taacgatatt
      601 attgaagcgc atcgtgaaca gattggcggc gataacccgg aacagtttga agtgctgaaa
      661 cagcagaaag aaattattga acagggcatt gatctgttta acaaaaaacc gaaacgcggc
      721 attcagtatc tgcaggaaca gggcatgctg ggcaccaccc cggaagatat tgcgcagttt
      781 ctgcatcagg aagaacgcct ggatagcacc caggtgggcg aatttctggg cgataacgat
      841 aaatttaaca aagaagtgat gtatgcgtat gtggatcagc atgattttag cggcaaagat
      901 tttgtgagcg cgctgcgcat gtttctggaa ggctttcgcc tgccgggcga agcgcagaaa
      961 attgatcgcc tgatggaaaa atttgcggcg cgctatctgg aatgcaacca gggccagacc
     1021 ctgtttgcga gcgcggatac cgcgtatgtg ctggcgtata gcattattat gctgaccacc
     1081 gatctgcata gcccgcaggt gaaaaacaaa atgaccaaag aacagtatat taaaatgaac
     1141 cgcggcatta acgatagcaa agatctgccg gaagaatatc tgagcgcgat ttataacgaa
     1201 attgcgggca aaaaaattag catgaaataa taaggatcca tgggtaccct cgagtgaaga
     1261 caagcttagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgagc
     1321 accaccacca ccaccactga gtccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1381 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1441 gggttttttg ctgaaaggag gaactatacc ggattggcga atgggacgcg ccctgtagcg
     1501 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1561 ccctagcgcc cgctcctttc gctttcttcc cttccttctc gccacgttcg ccggctttcc
     1621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt ggccatcgcc ctgatagacg
     1741 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1801 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat ttgccgattt
     1861 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1921 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctattgt
     1981 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     2041 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     2101 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     2161 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     2221 tcgtcaaaaa aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2281 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2341 catcaaaatc actcgcacaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2401 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2461 gaacactgcc agcgcatcaa caatttttca cctgaatcag gatattcttc taatacctgg
     2521 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     2581 aaatgcttga tggtcggaag aggcataaat tcgtcagcca gtttagtctg accatctcat
     2641 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2701 gcttcccata caatcgatag attgtcgcac ctgattgccg acattatcgc gagcccattt
     2761 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     2821 ccgttgaata tggctcataa caccccttgt attactgttt atgtagcaga cagttttatt
     2881 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gctgcaaaca
     3001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagcct
     3121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3241 gatagtaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3361 cacgcttccc gaaggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3421 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3481 cgccacctct gacttgagcg cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3601 tgttctttcc tgcgttatcc cctgatttgt ggataaccgt attaccgcct ttgagtgagc
     3661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3721 agagcgcctg atgcggtatt ttctccttac gcattgtgcg gtatttcaca ccgcaatggt
     3781 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     3841 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca aacccgctga cgcgccctga
     3901 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3961 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctggg taaagctcat
     4021 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     4081 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     4141 tttcctgttt ggtcactgat gcctccgtgt aagggggatt ctgttcatgg gggtaatgat
     4201 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     4261 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcggacca gagaaaaatc
     4321 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4381 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccggtttcca
     4441 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4501 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4561 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     4621 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     4681 gacgaagctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4741 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4801 cctacgagtt gcatataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4861 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4921 cctaatgagt gagctaactt aattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4981 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5041 tattgggcgc cagggtggtt tttcttttac cagtgagacg ggcaacagct gattgccctt
     5101 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5161 aaaatcctgt ttgatggtgg ttaacggcgg gatatacatg agctgtcttc ggtatcgtcg
     5221 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     5281 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tggaacgatg ccctcattca
     5341 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     5401 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacga gacgcgccga
     5461 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5521 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgctgg
     5581 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5641 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5701 tgcccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     5761 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     5821 ccagactgga gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5881 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     5941 agaaacgtgg ctggcctgtt caccacgcgg gaaacggtct gataagagac accggcatac
     6001 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6061 gggcgctatc atgccatacc gcgaaggttt tgcgccattc gatggtgtcc gggatctcga
     6121 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6181 agcaccgccg ccgcaaggaa tggtgcatg
//
