LOCUS       Exported                8309 bp ds-DNA     circular SYN 09-SEP-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8309)
  AUTHORS   Aleksandar Vjestica
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 9, 2019 from SnapGene 4.3.7
            https://www.snapgene.com

FEATURES             Location/Qualifiers
     source          1..8309
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    193..653
                     /label=3''region lys3
                     /note="3''region lys3"
     misc_feature    654..996
                     /label=5'region lys3
                     /note="5'region lys3"
     CDS             997..2103
                     /label=ORF lys3
                     /note="ORF lys3"
     misc_feature    2104..2470
                     /label=3'region lys3
                     /note="3'region lys3"
     misc_feature    2526..2981
                     /label=F1 ori
                     /note="F1 ori"
     primer_bind     3127..3144
                     /label=M13 forward
                     /note="M13 forward"
     primer_bind     3154..3173
                     /label=T7
                     /note="T7"
     misc_feature    3181..3185
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        3187..4174
                     /label=BiP promoter
                     /note="BiP promoter"
     CDS             4175..4249
                     /codon_start=1
                     /label=BiP Signal Sequence
                     /note="BiP Signal Sequence"
                     /translation="MKKFQLFSILSYFVALFLLPMAFAS"
     CDS             4274..4984
                     /codon_start=1
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSRLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4985..4999
                     /codon_start=1
                     /label=ADEL ER-retention signal
                     /translation="EADEL"
     misc_feature    5009..5391
                     /label=Ashbya gossypii TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5392..5784
                     /label=bsd ORF
                     /note="bsd ORF"
     misc_feature    5785..6017
                     /label=Ashbya gossypii TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6047..6066)
                     /label=T3
                     /note="T3"
     primer_bind     complement(6084..6104)
                     /label=M13 reverse
                     /note="M13 reverse"
     protein_bind    complement(6110..6132)
                     /label=lacO
                     /note="lacO"
     promoter        complement(6136..6166)
                     /label=lac promoter
                     /note="lac promoter"
     rep_origin      complement(6472..7100)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     CDS             complement(7249..8109)
                     /label=AmpR
                     /note="AmpR"
     promoter        complement(8110..8214)
                     /label=AmpR promoter
                     /note="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaccg gggccttacg aagttaaatt ataggcacag acatctttgg cattggaaag
      241 atttagtaac aatcataatg tagcacttct taaaaaatct aattttagaa caaaaaggct
      301 taaaaaagaa tggttccgct cttttaaaaa atagtaaaat gtacaatagc aaggttttcg
      361 taggtcgacg gacataacaa ggggaaagag aaataaagca tgtaaacgtc aataaaacat
      421 ctttgattct ggatcaatca tatgccaaat gaagaaaaat agcatttata acaacatgaa
      481 accatatttg agacctttta acgatgaaaa tttattaaat aaaagataaa aaagaaagaa
      541 accgttcaag ttcaatgaaa atttcgtcgt gaagtagaaa ccaagataat ggaaaggctt
      601 cgcaaagcca caaattttcc aatagtcatt ataactgaag attaaggaca gtatactagt
      661 aaaaatcaag tgaattgtgc ctaggatagg tagctaacgt gactgccact gaacctgtgt
      721 agcgtattag atccatcaag atcccttctg ctcatatcat tgctgatctg taaattgacc
      781 gcatcaaatt tgatctcatt tatatcgaga ttgatctata ttattttgcc aatttgcagt
      841 atataaactt atttgattaa gtttttggtc tccttcgcca cgtcttcatt ttgttagttt
      901 aaattttttt gactaactta gaaagggaag tttttaaaaa cgttgaaata ctttttcgat
      961 ttaaaaatat atatttaatt aattttctgt tccaaaatgg ttgctcctca tctttggtta
     1021 cgtgctgaaa ccaaaccttt agaagagcgc agtgctttga ctcctagaac tgccaagata
     1081 ctcgccgatg ctggatttca aattaccatt gagcgctctt cccaacgtgc tttcaaggac
     1141 aaagaatttg agcgccttgg tttccccatg gttcctgagg gctcatggcg acatgcacct
     1201 aaggacgcgt acattattgg tctcaaagag cttccggaaa atgataacag tcctttgaag
     1261 catactcata ttcaatttgc tcattgctac aaaaaccagg aaggttggcg tgaagtcttg
     1321 tcccgattcc ccgctggcaa tggtcttcta tatgatttgg aatttttgca agatgacaat
     1381 ggcagacgtg ttgccgcttt tggttaccac gctggttttg ctggatcagc tatctcatgt
     1441 ttagtttggg cacatcaatt attgcatcct aacaaacaat tccctgctat ccgtccgttc
     1501 cccaacgaaa aatctttggt tcgccatgtt gctcgtcaag tacgtttagc tttgaaaaag
     1561 aataacaacc agtatcctcg tattctggtt attggggctc ttggtcgttg tggtactgga
     1621 gcgtgcgatt tagcatcaaa gattggtatt cccttcgata acattcttcg ctgggacatt
     1681 aatgaaacta agaagggtgg tccctttacc gaaattacgg aatcagacat atttgttaac
     1741 tgcatttatc tgagcatgcc tattcccaaa ttctgtaccg tcgagtcgtt aaatgtacct
     1801 aaccgtaaat tgcgtgttgt ttgtgatgtt agctgtgaca ctaccaatcc taacaatcct
     1861 attccaattt ataatgtaaa cactacattt gatcatccta cagttgaggt taaaggtgtc
     1921 acaactcctc ctcctttaga ggtcatttct attgatcatc ttcctacttt attgccaagg
     1981 gaatcttccg aagcatttag tgaagctcta attcctagtc ttttggcttt gaaggacgtc
     2041 gacaatgctc ccgtttgggt cagagctaaa aaactttatg aaaccatggt tcaaaagctt
     2101 taaaataaat gttatttaat ttcaaagttc cgggtccttt atattgtttc accaaattat
     2161 tgaatattat tttctcaaaa attaacttta gaatacaatt tagacggcat tttcgatgct
     2221 ctggacattt tttccacgta cttgttaaat tgtagcggtg ttgcataata ctaccttttg
     2281 atttgttatt cacgttacaa tatatatttg actaatttaa atatgataat atacacattt
     2341 ctctgcaaac ctcctttata cttttcgccg gtcattgatt aataattaag aaacaggatt
     2401 tcatcctgca tttgccactt acctatatgg taaatataaa tcttgaaact ttccccaata
     2461 tttattgctt gctagggcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2521 tcaggaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcgcgccatt cgccattcag
     3001 gctgcgcaac tgttgggaag ggcgatcgcg gtgcgggcct cttcgctatt acgccagctg
     3061 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     3121 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     3181 ggtaccgtgc caagggaagg gtaggtgtcg ttaaggtctg aggtttggtg tcagtgggag
     3241 atctaaaata ttatggcaaa attttcggcg tcgcaacaat aatagtaaac agatgtaagg
     3301 actataccat tttgagatgt taaaatggtg agcaatgata caagtgatga tctatcatct
     3361 tttgctgatt gataaaaaaa gacaaaccct ttaattattt ttagctacat aattttctct
     3421 taatggatta atttggaaag tgaacgcaaa atattattta gtctaaacat gatcgtattt
     3481 gagatgatgc cttttgcaag tatgttatac tttatttaac catgactttt acaattcaac
     3541 caatgcgaca ctatcggaca gttgaagcca attttaatta ttagctatat attttagcgt
     3601 ttttccatta tatttgtagt taaaacagct atttataggc cattatcaaa tatatatata
     3661 attttttttt tttttttttt ctctttcatt tgggaatata gtgaaataac tctatataac
     3721 acaatatcaa aataaagatt tcccaccttc actgcttttg ttcttttcac aattattcat
     3781 ttcagaattt accaatttcg attttcattg tttacattca ttgctatgaa aacgtaaggt
     3841 ggtggcgcaa taggacttat cgaaatgtac agaactcact atagaattgt tgtgttgatg
     3901 agcttcaact gcattcttct ggaaagtact agtattaacg acgtgactgc tcctctcgtt
     3961 acttagctga tttctggtac gctattaaac tcatccaaaa ccaactattc tagtttggta
     4021 aatcttaatc aaaaactatt aaaacccgtt tactatttac ttaacaggtt gttttcaata
     4081 attgggaatt ggcttgtgcc tacgatctct tgtaattgaa ctacacatat aagcatttat
     4141 aagttggtaa tcttcaaatt cttgtttatt gaaaatgaag aagttccagc tatttagcat
     4201 tttaagctac tttgtagctt tattcctcct acctatggct tttgctagtg gatccggtgg
     4261 tcccggtggt atgtccaagg gtgaagagct atttactggg gttgtaccca ttttggtaga
     4321 actggacgga gatgtaaacg gacataaatt ctctgttaga ggtgagggcg aaggcgatgc
     4381 caccaatggt aaattgactc tgaagtttat atgcactacg ggtaaattac ctgttccttg
     4441 gccaacccta gtaacaactt tgacatatgg tgttcaatgt ttctcaagat acccagacca
     4501 tatgaaaagg catgatttct ttaaaagtgc tatgccagaa ggctacgtgc aagagagaac
     4561 tatctccttt aaggatgacg gtacgtataa aacacgagca gaagtgaaat tcgaagggga
     4621 tacactagtt aatcgcatcg aattaaaggg tatagacttt aaggaagatg gtaatattct
     4681 cggccataaa cttgagtata atttcaactc gcataatgtg tacattacag ctgacaaaca
     4741 aaagaacgga attaaagcga attttaaaat caggcacaac gtcgaagatg ggtctgttca
     4801 acttgccgat cattatcagc aaaacacccc tattggtgat ggtccagtct tgttacccga
     4861 taatcactac ttaagcacac agtctagatt gtcaaaagat ccgaatgaaa agcgtgatca
     4921 catggtttta ttggaatttg tcaccgctgc aggaataact cacggaatgg acgagcttta
     4981 taaggaggcc gatgaacttt aagagctcgt ttagcttgcc tcgtccccgc cgggtcaccc
     5041 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     5101 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     5161 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     5221 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     5281 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     5341 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgcctttg
     5401 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     5461 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5521 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5581 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5641 atcttgagcc cctgcggacg gtgtcgacag gtgcttctcg atctgcatcc tgggatcaaa
     5701 gcgatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5761 tctggttatg tgtgggaggg ctaatcagta ctgacaataa aaagattctt gttttcaaga
     5821 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     5881 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     5941 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     6001 actaacgccg ccatccacct gcaggtaatc acgtgccagc ttttgttccc tttagtgagg
     6061 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6121 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     6181 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6241 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6301 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6361 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6421 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6481 gctggcgttt ttccataggc tcggcccccc tgacgagcat cacaaaaatc gacgctcaag
     6541 tcagaggtgg cgaaacccga caggactata aagataccag gcgttccccc ctggaagctc
     6601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6661 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6901 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7321 gcccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7621 ccaacgatca aggcgagtta catgatcccc catgttgtga aaaaaagcgg ttagctcctt
     7681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7981 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8041 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8101 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8161 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8221 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8281 aaataggcgt atcacgaggc cctttcgtc
//
