LOCUS       Exported                8662 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 8662)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 31, 2021 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8662
                     /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             4144..8043
                     /codon_start=1
                     /label=FnCas12a
                     /label=Cpf1
                     /translation="MSIYQEFVNKYSLSKTLRFELIPQGKTLENIKARGLILDDEKRAK
                     DYKKAKQIIDKYHQFFIEEILSSVCISEDLLQNYSDVYFKLKKSDDDNLQKDFKSAKDT
                     IKKQISEYIKDSEKFKNLFNQNLIDAKKGQESDLILWLKQSKDNGIELFKANSDITDID
                     EALEIIKSFKGWTTYFKGFHENRKNVYSSNDIPTSIIYRIVDDNLPKFLENKAKYESLK
                     DKAPEAINYEQIKKDLAEELTFDIDYKTSEVNQRVFSLDEVFEIANFNNYLNQSGITKF
                     NTIIGGKFVNGENTKRKGINEYINLYSQQINDKTLKKYKMSVLFKQILSDTESKSFVID
                     KLEDDSDVVTTMQSFYEQIAAFKTVEEKSIKETLSLLFDDLKAQKLDLSKIYFKNDKSL
                     TDLSQQVFDDYSVIGTAVLEYITQQIAPKNLDNPSKKEQELIAKKTEKAKYLSLETIKL
                     ALEEFNKHRDIDKQCRFEEILANFAAIPMIFDEIAQNKDNLAQISIKYQNQGKKDLLQA
                     SAEDDVKAIKDLLDQTNNLLHKLKIFHISQSEDKANILDKDEHFYLVFEECYFELANIV
                     PLYNKIRNYITQKPYSDEKFKLNFENSTLANGWDKNKEPDNTAILFIKDDKYYLGVMNK
                     KNNKIFDDKAIKENKGEGYKKIVYKLLPGANKMLPKVFFSAKSIKFYNPSEDILRIRNH
                     STHTKNGSPQKGYEKFEFNIEDCRKFIDFYKQSISKHPEWKDFGFRFSDTQRYNSIDEF
                     YREVENQGYKLTFENISESYIDSVVNQGKLYLFQIYNKDFSAYSKGRPNLHTLYWKALF
                     DERNLQDVVYKLNGEAELFYRKQSIPKKITHPAKEAIANKNKDNPKKESVFEYDLIKDK
                     RFTEDKFFFHCPITINFKSSGANKFNDEINLLLKEKANDVHILSIARGERHLAYYTLVD
                     GKGNIIKQDTFNIIGNDRMKTNYHDKLAAIEKDRDSARKDWKKINNIKEMKEGYLSQVV
                     HEIAKLVIEYNAIVVFEDLNFGFKRGRFKVEKQVYQKLEKMLIEKLNYLVFKDNEFDKT
                     GGVLRAYQLTAPFETFKKMGKQTGIIYYVPAGFTSKICPVTGFVNQLYPKYESVSKSQE
                     FFSKFDKICYNLDKGYFEFSFDYKNFGDKAAKGKWTIASFGSRLINFRNSDKNHNWDTR
                     EVYPTKELEKLLKDYSIEYGHGECIKAAICGESDKKFFAKLTSVLNTILQMRNSKTGTE
                     LDYLISPVADVNGNFFDSRQAPKNMPQDADANGAYHIGLKGLMLLGRIKNNQEGKKLNL
                     VIKNEEYFEFVQNRNN"
     misc_difference 6892..6894
                     /label=D917A
     CDS             8044..8052
                     /codon_start=1
                     /label=linker
                     /translation="GSS"
     CDS             8053..8070
                     /codon_start=1
                     /label=sorting motif
                     /translation="LPETGG"
     CDS             8071..8088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      8212..8259
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(8625..8653)
                     /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 tttagttcca
     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 aagcggaaca 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 catatgtcaa tttatcaaga atttgttaat aaatatagtt taagtaaaac tctaagattt
     4201 gagttaatcc cacagggtaa aacacttgaa aacataaaag caagaggttt gattttagat
     4261 gatgagaaaa gagctaaaga ctacaaaaag gctaaacaaa taattgataa atatcatcag
     4321 ttttttatag aggagatatt aagttcggtt tgtattagcg aagatttatt acaaaactat
     4381 tctgatgttt attttaaact taaaaagagt gatgatgata atctacaaaa agattttaaa
     4441 agtgcaaaag atacgataaa gaaacaaata tctgaatata taaaggactc agagaaattt
     4501 aagaatttgt ttaatcaaaa ccttatcgat gctaaaaaag ggcaagagtc agatttaatt
     4561 ctatggctaa agcaatctaa ggataatggt atagaactat ttaaagccaa tagtgatatc
     4621 acagatatag atgaggcgtt agaaataatc aaatctttta aaggttggac aacttatttt
     4681 aagggttttc atgaaaatag aaaaaatgtt tatagtagca atgatattcc tacatctatt
     4741 atttatagga tagtagatga taatttgcct aaatttctag aaaataaagc taagtatgag
     4801 agtttaaaag acaaagctcc agaagctata aactatgaac aaattaaaaa agatttggca
     4861 gaagagctaa cctttgatat tgactacaaa acatctgaag ttaatcaaag agttttttca
     4921 cttgatgaag tttttgagat agcaaacttt aataattatc taaatcaaag tggtattact
     4981 aaatttaata ctattattgg tggtaaattt gtaaatggtg aaaatacaaa gagaaaaggt
     5041 ataaatgaat atataaatct atactcacag caaataaatg ataaaacact caaaaaatat
     5101 aaaatgagtg ttttatttaa gcaaatttta agtgatacag aatctaaatc ttttgtaatt
     5161 gataagttag aagatgatag tgatgtagtt acaacgatgc aaagttttta tgagcaaata
     5221 gcagctttta aaacagtaga agaaaaatct attaaagaaa cactatcttt attatttgat
     5281 gatttaaaag ctcaaaaact tgatttgagt aaaatttatt ttaaaaatga taaatctctt
     5341 actgatctat cacaacaagt ttttgatgat tatagtgtta ttggtacagc ggtactagaa
     5401 tatataactc aacaaatagc acctaaaaat cttgataacc ctagtaagaa agagcaagaa
     5461 ttaatagcca aaaaaactga aaaagcaaaa tacttatctc tagaaactat aaagcttgcc
     5521 ttagaagaat ttaataagca tagagatata gataaacagt gtaggtttga agaaatactt
     5581 gcaaactttg cggctattcc gatgatattt gatgaaatag ctcaaaacaa agacaatttg
     5641 gcacagatat ctatcaaata tcaaaatcaa ggtaaaaaag acctacttca agctagtgcg
     5701 gaagatgatg ttaaagctat caaggatctt ttagatcaaa ctaataatct cttacataaa
     5761 ctaaaaatat ttcatattag tcagtcagaa gataaggcaa atattttaga caaggatgag
     5821 catttttatc tagtatttga ggagtgctac tttgagctag cgaatatagt gcctctttat
     5881 aacaaaatta gaaactatat aactcaaaag ccatatagtg atgagaaatt taagctcaat
     5941 tttgagaact cgactttggc taatggttgg gataaaaata aagagcctga caatacggca
     6001 attttattta tcaaagatga taaatattat ctgggtgtga tgaataagaa aaataacaaa
     6061 atatttgatg ataaagctat caaagaaaat aaaggcgagg gttataaaaa aattgtttat
     6121 aaacttttac ctggcgcaaa taaaatgtta cctaaggttt tcttttctgc taaatctata
     6181 aaattttata atcctagtga agatatactt agaataagaa atcattccac acatacaaaa
     6241 aatggtagtc ctcaaaaagg atatgaaaaa tttgagttta atattgaaga ttgccgaaaa
     6301 tttatagatt tttataaaca gtctataagt aagcatccgg agtggaaaga ttttggattt
     6361 agattttctg atactcaaag atataattct atagatgaat tttatagaga agttgaaaat
     6421 caaggctaca aactaacttt tgaaaatata tcagagagct atattgatag cgtagttaat
     6481 cagggtaaat tgtacctatt ccaaatctat aataaagatt tttcagctta tagcaaaggg
     6541 cgaccaaatc tacatacttt atattggaaa gcgctgtttg atgagagaaa tcttcaagat
     6601 gtggtttata agctaaatgg tgaggcagag cttttttatc gtaaacaatc aatacctaaa
     6661 aaaatcactc acccagctaa agaggcaata gctaataaaa acaaagataa tcctaaaaaa
     6721 gagagtgttt ttgaatatga tttaatcaaa gataaacgct ttactgaaga taagtttttc
     6781 tttcactgtc ctattacaat caattttaaa tctagtggag ctaataagtt taatgatgaa
     6841 atcaatttat tgctaaaaga aaaagcaaat gatgttcata tattaagtat agctagaggt
     6901 gaaagacatt tagcttacta tactttggta gatggtaaag gcaatatcat caaacaagat
     6961 actttcaaca tcattggtaa tgatagaatg aaaacaaact accatgataa gcttgctgca
     7021 atagagaaag atagggattc agctaggaaa gactggaaaa agataaataa catcaaagag
     7081 atgaaagagg gctatctatc tcaggtagtt catgaaatag ctaagctagt tatagagtat
     7141 aatgctattg tggtttttga ggatttaaat tttggattta aaagagggcg tttcaaggta
     7201 gagaagcagg tctatcaaaa gttagaaaaa atgctaattg agaaactaaa ctatctagtt
     7261 ttcaaagata atgagtttga taaaactggg ggagtgctta gagcttatca gctaacagca
     7321 ccttttgaga cttttaaaaa gatgggtaaa caaacaggta ttatctacta tgtaccagct
     7381 ggttttactt caaaaatttg tcctgtaact ggttttgtaa atcagttata tcctaagtat
     7441 gaaagtgtca gcaaatctca agagttcttt agtaagtttg acaagatttg ttataacctt
     7501 gataagggct attttgagtt tagttttgat tataaaaact ttggtgacaa ggctgccaaa
     7561 ggcaagtgga ctatagctag ctttgggagt agattgatta actttagaaa ttcagataaa
     7621 aatcataatt gggatactcg agaagtttat ccaactaaag agttggagaa attgctaaaa
     7681 gattattcta tcgaatatgg gcatggcgaa tgtatcaaag cagctatttg cggtgagagc
     7741 gacaaaaagt tttttgctaa gctaactagt gtcctaaata ctatcttaca aatgcgtaac
     7801 tcaaaaacag gtactgagtt agattatcta atttcaccag tagcagatgt aaatggcaat
     7861 ttctttgatt cgcgacaggc gccaaaaaat atgcctcaag atgctgatgc caatggtgct
     7921 tatcatattg ggctaaaagg tctgatgcta ctaggtagga tcaaaaataa tcaagagggc
     7981 aaaaaactca atttggttat caaaaatgaa gagtattttg agttcgtgca gaataggaat
     8041 aacggttctt ctctgccaga aacgggaggc catcaccatc atcaccacta acggatccgc
     8101 gatcgtggcg cgccacctgg tggccggccg gtaccacgcg tgcgcgctga tccggctgct
     8161 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     8221 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     8281 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
     8341 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     8401 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     8461 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     8521 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     8581 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     8641 gataagctgt caaacatgag aa
//
