LOCUS       MZB224        7082 bp DNA     circular SYN 15-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  AUTHORS   UNC Chapel Hill
  TITLE     Direct Submission
  JOURNAL   Exported Sep 15, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        208..226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     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=SUMO (Smt3)
     misc_feature    609..630
                     /label=SUMO HiFi Rev
     CDS             631..1521
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1537..1560
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    1651..1653
                     /label=T31N
     misc_feature    2101..2129
                     /label=SUMO HiFi For
     CDS             2193..2210
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2262..2280)
                     /label=T7-Term
     terminator      2276..2323
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2359..2811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3832..4415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4600..4738
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        complement(6906..6982)
                     /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 atggcagaaa tcggtactgg ctttccattc
      661 gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc
      721 gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac
      781 atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc
      841 aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc
      901 ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct
      961 ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag
     1021 ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag
     1081 gccttccgca ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag
     1141 ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc
     1201 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
     1261 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
     1321 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
     1381 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
     1441 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg
     1501 tcgacgctcg agatttccgg cgagccaacc actggcctgg aagtgctgtt tcagggcccg
     1561 ggtaatatat tcgccaactt gttcaagggc ttgttcggta aaaaagagat gcgcattttg
     1621 atggtcggtc tggacgctgc cggcaaaacg aacatcctgt ataagctgaa attaggggaa
     1681 atcgtgacta ccattcctac aatcggcttt aacgtagaaa cagttgaata taagaatatc
     1741 agttttactg tatgggatgt agggggtcag gataagatca gacctttatg gagacactat
     1801 ttccaaaata cgcagggact gatctttgta gtagactcta atgaccgtga gcgggtgaat
     1861 gaagcccgcg aagaacttat gcgcatgctt gcagaagacg agctgagaga tgcggtcctg
     1921 ttggtgttcg ccaataaaca agatttaccg aacgccatga acgctgcaga aataactgac
     1981 aaattgggac ttcactccct gcggcataga aactggtaca ttcaggcaac gtgcgcaacg
     2041 agcggcgacg ggctgtacga gggtttggac tggcttagca accaactgcg gaatcaaaaa
     2101 taataaggat ccatgggtac cctcgagtga agacaagctt aggtatttat tcggcgcaaa
     2161 gtgcgtcggg tgatgctgcc aacttagtcg agcaccacca ccaccaccac tgagtccggc
     2221 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2281 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2341 accggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2401 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2461 tcccttcctt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2521 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2581 tggttcacgt agtggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2701 tattcttttg atttataagg gatttgccga tttcggccta ttggttaaaa aatgagctga
     2761 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2821 ttttcgggga aatgtgcgcg gaacccctat tgtttatttt tctaaataca ttcaaatatg
     2881 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2941 attcatatca ggattatcaa taccatattt ttgaaaagcc gtttctgtaa tgaaggagaa
     3001 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     3061 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaaaaggtta tcaagtgaga
     3121 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     3181 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca caaccaaacc
     3241 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3301 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatttt
     3361 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3421 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3481 aattcgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     3541 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     3601 cacctgattg ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     3661 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     3721 tgtattactg tttatgtagc agacagtttt attgttcatg accaaaatcc cttaacgtga
     3781 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3841 tttttttctg cgcgtaatct gctgctgcaa acaaaaaaac caccgctacc agcggtggtt
     3901 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3961 cagataccaa atactgtcct tctagtgtag cctagttagg ccaccacttc aagaactctg
     4021 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4081 ataagtcgtg tcttaccggg ttggactcaa gacgatagta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggag aaaggcggac
     4261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgcgatttt
     4381 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4441 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4501 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4561 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4621 tacgcattgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     4681 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4741 ccgacacccg ccaaacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4801 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4861 ccgaaacgcg cgaggcagct gggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4921 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4981 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     5041 tgtaaggggg attctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     5101 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     5161 ggcggtatgg atgcggcgga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     5221 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5281 ataatggtgc agggcgctga cttccggttt ccagacttta cgaaacacgg aaaccgaaga
     5341 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     5401 cgcgtatcgg tgattcattc tgctaaccag taagcaaccc cgccagccta gccgggtcct
     5461 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     5521 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag cttgagcgag ggcgtgcaag
     5581 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5641 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatata aagaagacag
     5701 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     5761 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttaattaat
     5821 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5881 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     5941 taccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     6001 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     6061 cgggatatac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     6121 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     6181 accagcatcg cagtggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     6241 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     6301 atttatgcca gccagccaga cgagacgcgc cgagacagaa cttaatgggc ccgctaacag
     6361 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     6421 atgggagaaa ataatactgt tgatgggtgc tggtcagaga catcaagaaa taacgccgga
     6481 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     6541 atcagcccac tgacgcgttg cgcgagaaga ttgtgcccgc cgctttacag gcttcgacgc
     6601 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     6661 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggagtggcaa cgccaatcag
     6721 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6781 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct gttcaccacg
     6841 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     6901 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaagg
     6961 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     7021 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     7081 tg
//
