LOCUS       Exported                9094 bp ds-DNA     circular SYN 18-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9094)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9094
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             296..1237
                     /codon_start=1
                     /label=Spectinomycin adenyltransferase
                     /label=Spectinomycin adenyltransferase
                     /translation="MACYDCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPK
                     TKLNIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTV
                     TVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQ
                     RNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTL
                     WNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAY
                     LGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     Replication_ori complement(1546..4331)
                     /direction=LEFT
                     /label=pVS1ori short
                     /note="/vntifkey=21"
     misc_feature    4403..4440
                     /label=Left border
                     /label=nonstandard type: Region
     promoter        4995..5340
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             complement(5394..5699)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(6041..6700)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(6754..6784)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6944..6970
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6974..7000
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7004..7030
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7037..7993
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     terminator      8001..8253
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    complement(8314..8358)
                     /label=Right border
                     /label=nonstandard type: Region
     Replication_ori 8461
                     /label=ORI
                     /note=" RNaseH cleavage point "
                     /note="/vntifkey=33"
     Replication_ori complement(8565..8845)
                     /direction=LEFT
                     /label=pBR322 origin of replication
ORIGIN
        1 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
       61 atcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
      121 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaattcggg cacgaaccca
      181 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
      241 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
      301 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
      361 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
      421 tcgccctaaa acaaagttaa acatcatggg ggaagcggtg atcgccgaag tatcgactca
      481 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
      541 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
      601 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
      661 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
      721 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
      781 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
      841 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
      901 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
      961 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1021 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1081 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1141 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1201 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctagctagaa attcgttcaa
     1261 gccgacgccg cttcgcggcg cggcttaact caagcgttag atgcactaag cacataattg
     1321 ctcacagcca aactatcagg tcaagtctgc ttttattatt tttaagcgtg cataataagc
     1381 cctacacaaa ttgggagata tatcatgcat gaccaaaatc ccttaacgtg agttttcgtt
     1441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tctgccattt ttggggtgag
     1501 gccgttcgcg gccgaggggc gcagcccctg gggggatggg agctcggtgc cttgatgtgg
     1561 gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg
     1621 taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg
     1681 tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc
     1741 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
     1801 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
     1861 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
     1921 cccaatgtac gtgctatcca caggaaagaa gctttttcga cctttttccc ctgctagggc
     1981 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
     2041 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
     2101 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
     2161 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
     2221 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
     2281 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
     2341 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
     2401 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
     2461 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
     2521 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
     2581 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
     2641 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
     2701 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
     2761 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
     2821 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
     2881 aatcgacgac gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
     2941 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
     3001 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
     3061 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
     3121 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
     3181 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
     3241 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
     3301 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
     3361 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
     3421 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
     3481 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
     3541 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
     3601 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
     3661 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
     3721 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
     3781 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
     3841 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
     3901 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
     3961 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
     4021 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
     4081 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
     4141 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     4201 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     4261 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     4321 gcagccgcac ggctgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     4381 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
     4441 gacacttaat aacacattgc ggacgttttt aatgtactgg ggtggattac ggaagacaat
     4501 acggcggaga ttagcctttt caatttcaga aagaatgcta acccacagat ggttagagag
     4561 gcttacgcag cacgactcat caagacgatc tacccgagca ataatctcca ggaaatcaaa
     4621 taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg catcaagaac
     4681 acagagaaag atatatttct caagatcaga agtactattc cagtatggac gattcaaggc
     4741 ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt agttcccact
     4801 gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact cgccgtaaag
     4861 actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa aatcttcgtc
     4921 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     4981 gagcaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     5041 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     5101 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     5161 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaacacgt tccaaccacg
     5221 acttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     5281 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagagaaca
     5341 cgcacctgag acggtcgacc tgcagactgg ctgtgtataa gggagcctga catttatatt
     5401 ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct gagatcagcc
     5461 acttcttccc cgataacgga aaccggcaca ctggccatat cggtggtcat catgcgccag
     5521 ctttcatccc cgatatgcac caccgggtaa agttcacggg ggactttatc tgacagcaga
     5581 cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat atcactctgt
     5641 acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt aaactgcatt
     5701 tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc gggcgacctc
     5761 agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac gggcttcatt
     5821 ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt gagcaactga
     5881 tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct ctttttgaca
     5941 tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg cgcaaatacg
     6001 catactgtta tctggctttt agtaagccgg atcctctaga ttacgccccg ccctgccact
     6061 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     6121 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     6181 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     6241 tgaaactcac ccagggattg gctgacacga aaaacatatt ctcaataaac cctttaggga
     6301 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     6361 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     6421 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     6481 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     6541 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     6601 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     6661 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     6721 aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa gcataaagtg
     6781 taaagcctgg ggtgcctaat gcggcccctc gaggggccgc catagtgact ggatatgttg
     6841 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     6901 tatatcattt tacgtttctc gttcagcttt cgtctctaag gcatacccgt atgatgttcc
     6961 ggattacgct ggctatccct acgacgtgcc cgactatgcc gggtacccct atgacgtccc
     7021 agactacgca gctagcaaag acaatactgt gcctctgaag ctgatcgctc tcctggctaa
     7081 tggcgagttc catagtggcg aacagctggg agaaaccctg ggcatgtcca gggccgctat
     7141 caacaagcac attcagactc tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa
     7201 gggctactct ctgcccgagc ctatcccgct gctgaacgct aaacagattc tgggacagct
     7261 ggacggcggg agcgtggcag tcctgcctgt ggtagactcc accaatcagt acctgctgga
     7321 tcgaatcggc gagctgaaga gtggggatgc ttgcattgca gaatatcagc aggcagggag
     7381 aggaagcaga gggaggaaat ggttctctcc ttttggagct aacctgtacc tgagtatgtt
     7441 ttggcgcctg aagcggggac cagcagcaat cggcctgggc ccggtcatcg gaattgtcat
     7501 ggcagaagcg ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc ccaatgacct
     7561 gtatctgcag gatagaaagc tggcaggcat cctggtggag ctggccggaa taacaggcga
     7621 tgctgcacag atcgtcattg gcgccgggat taacgtggct atgaggcgcg tggaggaaag
     7681 cgtggtcaat cagggctgga tcacactgca ggaagcaggg attaacctgg acaggaatac
     7741 tctggccgct acgctgatcc gagagctgcg ggcagccctg gaactgttcg agcaggaagg
     7801 cctggctcca tatctgccac ggtgggagaa gctggataac ttcatcaata gacccgtgaa
     7861 gctgatcatt ggggacaaag agattttcgg gattagccgg gggattgata aacagggagc
     7921 cctgctgctg gaacaggacg gagttatcaa accctggatg ggcggagaaa tcagtctgcg
     7981 gtctgccgaa aagtagaatc gatcgttcaa acatttggca ataaagtttc ttaagattga
     8041 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     8101 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     8161 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     8221 tatcgcgcgc ggtgtcatct atgttactag atctgagtga cctaagctag cctagtgtcg
     8281 ggcttgtctt ctctggttta aactatcagt gtttgacagg atatattggc gggtaaacct
     8341 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     8401 cgttcgtcca tttgtatgtg ccaaccacag gaaccgtaaa aaggccgcgt tgctggcgtt
     8461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8881 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     8941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     9001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     9061 gatcttttct acggggtctg acgctcagtg gaac
//
