LOCUS       Exported                7704 bp ds-DNA     circular SYN 31-JAN-2017
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...1405975899002.105
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7704)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2018 from SnapGene 4.1.3
            http://www.snapgene.com
COMMENT     f1:ctcactaaagggaacaaaagctggagctccgcaaagaagtttaataatcatattac
            r1:gcgtgacataactaattacatgactcgagctattaCTTATCGTCGTCATCCTTGTAGTC
            Sac, XhoI
FEATURES             Location/Qualifiers
     source          1..7704
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..1107
                     /codon_start=1
                     /gene="LEU2"
                     /label=LEU2
                     /label=LEU2(1)
                     /note="Orf frame 3"
                     /translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
                     ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKEL
                     QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
                     EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
                     NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
                     IRTGDLGGSNSTTEVGDAVAEEVKKILA"
     gene            1..1107
                     /gene="LEU2"
                     /label=LEU2
     primer_bind     1719..1741
                     /label=pGEX_3_primer
     misc_feature    complement(1834..2352)
                     /label=CEN6_ARS4
     CDS             2484..3344
                     /codon_start=1
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /label=Ampicillin(1)
                     /note="Orf frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2484..3344
                     /gene="Ampicillin"
                     /label=Ampicillin
     CDS             2682..3341
                     /label=AmpR
     rep_origin      3439..4121
                     /label=ColE1 origin
     rep_origin      3499..4118
                     /label=pBR322_origin
     misc_binding    4461..4483
                     /label=LacO
     primer_bind     4470..4492
                     /label=M13_pUC_rev_primer
     primer_bind     4489..4509
                     /label=M13-rev
     primer_bind     4491..4509
                     /label=M13_reverse_primer
     primer_bind     4527..4546
                     /label=T3
     misc_feature    4563..5202
                     /label=Gal 1 Promoter
     CDS             5260..5262
                     /codon_start=1
                     /label=New Feature(1)
                     /translation="M"
     CDS             5263..6033
                     /codon_start=1
                     /label=BirA (humanized)
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    5266..5268
                     /label=Q65P
     misc_feature    5332..5334
                     /label=I87V
     misc_feature    5416..5418
                     /label=G115G
     misc_feature    5425..5427
                     /label=R118S
     misc_feature    5491..5493
                     /label=E141K
     misc_difference 5494..5496
                     /label=Q142R
     misc_feature    5518..5520
                     /label=S150G
     misc_feature    5521..5523
                     /label=L151P
     misc_feature    5548..5550
                     /label=V160A
     misc_feature    5644..5646
                     /label=T192A
     misc_feature    5650..5652
                     /label=K194I
     misc_feature    5695..5697
                     /label=M209V
     misc_structure  5941..5943
                     /label=G291G
     misc_feature    5983..5985
                     /label=I305V
     misc_feature    6034..6039
                     /label=NheI
     misc_feature    6040..6081
                     /label=V5 tag
     misc_feature    6088
                     /label=Geneious name: CHIMERA Pcr product - Fragment 2 ex
                     /note="Geneious type: Concatenated sequence"
     terminator      6094..6341
                     /label=CYC1_terminator
                     /note="CYC1_terminator;26to274"
     primer_bind     complement(6359..6379)
                     /label=T7
     primer_bind     complement(6388..6405)
                     /label=M13-fwd
     primer_bind     complement(6388..6404)
                     /label=M13_forward20_primer
     misc_feature    6390..6533
                     /label=lacZ_a
     primer_bind     complement(6397..6419)
                     /label=M13_pUC_fwd_primer
     CDS             6476..6544
                     /label=LacZ alpha
     rep_origin      6545..7000
                     /label=M13 origin
     rep_origin      6562..6868
                     /label=F1 ori
     rep_origin      6562..6868
                     /label=f1_origin
ORIGIN
        1 atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt
       61 caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat
      121 gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc
      181 ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct
      241 gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc
      301 cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt
      361 ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga
      421 gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct
      481 tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc
      541 atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg
      601 gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca
      661 ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc
      721 cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc
      781 tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac
      841 aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag
      901 aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg
      961 aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt
     1021 atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc
     1081 gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata
     1141 aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca
     1201 tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag
     1261 gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa
     1321 gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt
     1381 aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa
     1441 aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa
     1501 taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag
     1561 aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc
     1621 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     1681 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     1741 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     1801 ataggttaat gtcatgataa taatggtttc ttaggacgga tcgcttgcct gtaacttaca
     1861 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     1921 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     1981 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     2041 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     2101 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     2161 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     2221 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     2281 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     2341 atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3181 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3661 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4141 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4201 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4261 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     4321 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4381 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     4441 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     4501 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     4561 tccgcaaaga agtttaataa tcatattaca tggcattacc accatataca tatccatata
     4621 catatccata tctaatctta cttatatgtt gtggaaatgt aaagagcccc attatcttag
     4681 cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt
     4741 gaagtacgga ttagaagccg ccgagcgggt gacagccctc cgaaggaaga ctctcctccg
     4801 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     4861 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     4921 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa
     4981 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     5041 tctattaaca gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca
     5101 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa
     5161 tatacctcta tactttaacg tcaaggagaa aaaaccccgg atcgaattct acttcataca
     5221 ttttcaatta agatggctcg ggatccaccg gtcgccacca tgatcccgct gctgaacgct
     5281 aaacagattc tgggacagct ggacggcggg agcgtggcag tcctgcctgt ggtcgactcc
     5341 accaatcagt acctgctgga tcgaatcggc gagctgaaga gtggggatgc ttgcattgca
     5401 gaatatcagc aggcagggag aggaagcaga gggaggaaat ggttctctcc ttttggagct
     5461 aacctgtacc tgagtatgtt ttggcgcctg aagcggggac cagcagcaat cggcctgggc
     5521 ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc tgggagcaga caaggtgcga
     5581 gtcaaatggc ccaatgacct gtatctgcag gatagaaagc tggcaggcat cctggtggag
     5641 ctggccggaa taacaggcga tgctgcacag atcgtcattg gcgccgggat taacgtggct
     5701 atgaggcgcg tggaggaaag cgtggtcaat cagggctgga tcacactgca ggaagcaggg
     5761 attaacctgg acaggaatac tctggccgct atgctgatcc gagagctgcg ggcagccctg
     5821 gaactgttcg agcaggaagg cctggctcca tatctgtcac ggtgggagaa gctggataac
     5881 ttcatcaata gacccgtgaa gctgatcatt ggggacaaag agattttcgg gattagccgg
     5941 gggattgata aacagggagc cctgctgctg gaacaggacg gagttatcaa accctggatg
     6001 ggcggagaaa tcagtctgcg gtctgccgaa aaggctagcg gcaagcccat ccccaacccc
     6061 ctgctgggcc tggacagcac ctaatagctc gagtcatgta attagttatg tcacgcttac
     6121 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     6181 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     6241 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     6301 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     6361 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     6421 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6481 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6541 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6601 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6661 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6721 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6781 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6841 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6901 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6961 tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc
     7021 ttacgcatct gtgcggtatt tcacaccgca tatcgacggt cgaggagaac ttctagtata
     7081 tccacatacc taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc
     7141 cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt
     7201 atatttatag gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg
     7261 tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta
     7321 tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc aacataacga
     7381 gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat
     7441 ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag
     7501 gtgagaggcc ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg
     7561 catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg
     7621 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat
     7681 atatttcaag gatataccat tcta
//
