LOCUS       Exported                9251 bp ds-DNA     circular SYN 31-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 31, 2021 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9251
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             209..1069
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2014..2153
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2255..2446)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4007..4025
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4114..4136
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4156..4185
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4195..5295
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             5350..5370
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5377..8658
                     /codon_start=1
                     /label=Cas12i1
                     /translation="MSNKEKNASETRKAYTTKMIPRSHDRMKLLGNFMDYLMDGTPIFF
                     ELWNQFGGGIDRDIISGTANKDKISDDLLLAVNWFKVMPINSKPQGVSPSNLANLFQQY
                     SGSEPDIQAQEYFASNFDTEKHQWKDMRVEYERLLAELQLSRSDMHHDLKLMYKEKCIG
                     LSLSTAHYITSVMFGTGAKNNRQTKHQFYSKVIQLLEESTQINSVEQLASIILKAGDCD
                     SYRKLRIRCSRKGATPSILKIVQDYELGTNHDDEVNVPSLIANLKEKLGRFEYECEWKC
                     MEKIKAFLASKVGPYYLGSYSAMLENALSPIKGMTTKNCKFVLKQIDAKNDIKYENEPF
                     GKIVEGFFDSPYFESDTNVKWVLHPHHIGESNIKTLWEDLNAIHSKYEEDIASLSEDKK
                     EKRIKVYQGDVCQTINTYCEEVGKEAKTPLVQLLRYLYSRKDDIAVDKIIDGITFLSKK
                     HKVEKQKINPVIQKYPSFNFGNNSKLLGKIISPKDKLKHNLKCNRNQVDNYIWIEIKVL
                     NTKTMRWEKHHYALSSTRFLEEVYYPATSENPPDALAARFRTKTNGYEGKPALSAEQIE
                     QIRSAPVGLRKVKKRQMRLEAARQQNLLPRYTWGKDFNINICKRGNNFEVTLATKVKKK
                     KEKNYKVVLGYDANIVRKNTYAAIEAHANGDGVIDYNDLPVKPIESGFVTVESQVRDKS
                     YDQLSYNGVKLLYCKPHVESRRSFLEKYRNGTMKDNRGNNIQIDFMKDFEAIADDETSL
                     YYFNMKYCKLLQSSIRNHSSQAKEYREEIFELLRDGKLSVLKLSSLSNLSFVMFKVAKS
                     LIGTYFGHLLKKPKNSKSDVKAPPITDEDKQKADPEMFALRLALEEKRLNKVKSKKEVI
                     ANKIVAKALELRDKYGPVLIKGENISDTTKKGKKSSTNSFLMDWLARGVANKVKEMVMM
                     HQGLEFVEVNPNFTSHQDPFVHKNPENTFRARYSRCTPSELTEKNRKEILSFLSDKPSK
                     RPTNAYYNEGAMAFLATYGLKKNDVLGVSLEKFKQIMANILHQRSEDQLLFPSRGGMFY
                     LATYKLDADATSVNWNGKQFWVCNADLVAAYNVGLVDIQKDFKKK"
     terminator      8801..8848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(9214..9242)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     2101 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2281 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     2341 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     2401 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     2461 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     2521 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     2581 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     2641 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     2701 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     2761 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     2821 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     2881 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
     2941 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
     3001 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
     3061 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
     3121 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
     3181 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
     3241 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
     3301 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
     3361 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
     3421 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     3481 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     3541 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     3601 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     3661 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac gctctccctt
     3721 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     3781 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     3841 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3901 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3961 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     4021 atagggagac cacaacggtt tccctctagt gccggctccg gagagctctt taattaagcg
     4081 gccgccctgc aggactcgag ttctagaaat aattttgttt aactttaaga aggagatata
     4141 catatgaaat cttctcacca tcaccatcac catcaccatc accatggttc ttctatgaaa
     4201 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
     4261 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
     4321 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc
     4381 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
     4441 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
     4501 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
     4561 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
     4621 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
     4681 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
     4741 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
     4801 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
     4861 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
     4921 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
     4981 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa
     5041 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
     5101 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccactatg
     5161 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
     5221 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
     5281 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc
     5341 gggatcgagg aaaacctgta cttccaatcc aatgcaatga gcaacaaaga aaagaacgcg
     5401 agcgaaaccc gtaaggcgta caccaccaaa atgatcccgc gtagccacga ccgtatgaag
     5461 ctgctgggca actttatgga ctatctgatg gatggcaccc cgattttctt tgaactgtgg
     5521 aaccagttcg gtggcggtat cgaccgtgat atcattagcg gcaccgcgaa caaagataag
     5581 attagcgacg atctgctgct ggcggttaac tggtttaaag tgatgccgat caacagcaaa
     5641 ccgcaaggcg tgagcccgag caacctggcg aacctgttcc agcaatacag cggtagcgaa
     5701 ccggacatcc aggcgcaaga gtatttcgcg agcaactttg acaccgaaaa gcaccagtgg
     5761 aaagatatgc gtgttgagta cgaacgtctg ctggcggagc tgcaactgag ccgtagcgac
     5821 atgcaccatg atctgaagct gatgtacaaa gaaaagtgca ttggcctgag cctgagcacc
     5881 gcgcactata tcaccagcgt gatgtttggc accggtgcga agaacaaccg tcagaccaaa
     5941 catcaattct atagcaaagt gattcagctg ctggaggaaa gcacccagat caacagcgtt
     6001 gagcaactgg cgagcatcat tctgaaagcg ggtgactgcg atagctaccg taaactgcgt
     6061 atccgttgca gccgtaaggg tgcgaccccg agcattctga aaatcgtgca agactatgaa
     6121 ctgggcacca accacgacga tgaggtgaac gttccgagcc tgattgcgaa cctgaaagaa
     6181 aagctgggcc gttttgagta cgaatgcgag tggaagtgca tggagaaaat caaggcgttc
     6241 ctggcgagca aagttggccc gtactatctg ggtagctata gcgcgatgct ggagaacgcg
     6301 ctgagcccga ttaagggtat gaccaccaag aactgcaaat tcgtgctgaa acagattgac
     6361 gcgaagaacg atatcaaata cgaaaacgag ccgtttggca agatcgttga aggtttcttt
     6421 gacagcccgt atttcgagag cgataccaac gtgaaatggg ttctgcaccc gcaccatatt
     6481 ggcgaaagca acatcaagac cctgtgggag gacctgaacg cgattcatag caaatacgag
     6541 gaagacatcg cgagcctgag cgaagataag aaagagaagc gtatcaaggt ttaccagggc
     6601 gatgtgtgcc aaaccatcaa cacctattgc gaggaagtgg gcaaggaagc gaaaaccccg
     6661 ctggttcagc tgctgcgtta cctgtatagc cgtaaagacg atattgcggt ggacaagatc
     6721 attgatggta tcaccttcct gagcaagaaa cacaaggtgg agaaacagaa gattaacccg
     6781 gttatccaaa aatacccgag cttcaacttt ggcaacaaca gcaagctgct gggtaaaatc
     6841 attagcccga aagacaagct gaaacataac ctgaagtgca accgtaacca agtggataac
     6901 tacatctgga ttgaaatcaa ggttctgaac accaagacca tgcgttggga gaaacaccat
     6961 tatgcgctga gcagcacccg ttttctggag gaagtttact atccggcgac cagcgaaaac
     7021 ccgccggatg cgctggcggc gcgtttccgt accaaaacca acggttatga gggcaagccg
     7081 gcgctgagcg cggaacagat tgagcaaatc cgtagcgcgc cggttggcct gcgtaaagtg
     7141 aagaaacgtc agatgcgtct ggaagcggcg cgtcagcaaa acctgctgcc gcgttatacc
     7201 tggggcaagg attttaacat taacatctgc aaacgtggta acaacttcga agttaccctg
     7261 gcgaccaagg tgaagaaaaa gaaagagaag aactacaaag tggttctggg ttatgacgcg
     7321 aacattgtgc gtaaaaacac ctatgcggcg attgaggcgc atgcgaacgg tgatggtgtg
     7381 attgactata acgatctgcc ggttaagccg atcgaaagcg gctttgtgac cgttgagagc
     7441 caggttcgtg acaaaagcta cgatcaactg agctataacg gtgtgaagct gctgtactgc
     7501 aaaccgcatg ttgaaagccg tcgtagcttc ctggagaaat atcgtaacgg caccatgaag
     7561 gacaaccgtg gtaacaacat tcagatcgac ttcatgaagg attttgaagc gatcgcggac
     7621 gatgaaacca gcctgtacta ttttaacatg aagtactgca aactgctgca gagcagcatt
     7681 cgtaaccaca gcagccaagc gaaagaatat cgtgaggaaa tcttcgagct gctgcgtgat
     7741 ggcaagctga gcgtgctgaa actgagcagc ctgagcaacc tgagcttcgt gatgtttaag
     7801 gttgcgaaaa gcctgattgg cacctacttt ggtcatctgc tgaagaaacc gaagaacagc
     7861 aaaagcgacg ttaaggctcc gccgatcacc gacgaagata agcagaaagc ggacccggag
     7921 atgttcgcgc tgcgtctggc gctggaggaa aaacgtctga acaaggttaa aagcaagaaa
     7981 gaagtgattg cgaacaagat cgttgcgaaa gcgctggagc tgcgtgacaa atacggcccg
     8041 gtgctgatta agggcgagaa catcagcgat accaccaaga aaggtaagaa aagcagcacc
     8101 aacagcttcc tgatggactg gctggcgcgt ggcgtggcga acaaggttaa agaaatggtg
     8161 atgatgcacc agggtctgga gttcgttgag gtgaacccga actttaccag ccaccaagat
     8221 ccgttcgttc ataagaaccc ggagaacacc tttcgtgcgc gttacagccg ttgcaccccg
     8281 agcgaactga ccgagaagaa ccgtaaagaa atcctgagct tcctgagcga caagccgagc
     8341 aaacgtccga ccaacgcgta ctataacgag ggtgcgatgg cgtttctggc gacctatggc
     8401 ctgaagaaaa acgacgtgct gggtgttagc ctggaaaagt tcaaacagat tatggcgaac
     8461 atcctgcatc agcgtagcga ggatcaactg ctgtttccga gccgtggcgg tatgttctac
     8521 ctggcgacct ataagctgga cgcggatgcg accagcgtta actggaacgg caaacagttt
     8581 tgggtgtgca acgcggacct ggttgcggcg tacaacgtgg gtctggttga catccaaaaa
     8641 gatttcaaga aaaagtaata ataacattgg aagtggataa cggatccgcg atcgcggcgc
     8701 gccacctggt ggccggccgg taccacgcgt gcgcgctgat ccggctgcta acaaagcccg
     8761 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     8821 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca
     8881 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca
     8941 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt
     9001 gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga
     9061 cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     9121 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     9181 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
     9241 aaacatgaga a
//
