LOCUS       Exported                7890 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 7890)
  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..7890
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     gene            1..1107
                     /gene="LEU2"
                     /label=LEU2
     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"
     primer_bind     1719..1741
                     /label=pGEX_3_primer
     misc_feature    complement(1834..2352)
                     /label=CEN6_ARS4
     gene            2484..3344
                     /gene="Ampicillin"
                     /label=Ampicillin
     CDS             2484..3344
                     /codon_start=1
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /label=Ampicillin(1)
                     /note="Orf frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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..6219
                     /codon_start=1
                     /label=BirA (humanized)
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    5452..5454
                     /label=Q65P
     misc_feature    5518..5520
                     /label=I87V
     misc_feature    5602..5604
                     /label=G115G
     misc_feature    5611..5613
                     /label=R118S
     misc_feature    5677..5679
                     /label=E141K
     misc_difference 5680..5682
                     /label=Q142R
     misc_feature    5704..5706
                     /label=S150G
     misc_feature    5707..5709
                     /label=L151P
     misc_feature    5734..5736
                     /label=V160A
     misc_feature    5830..5832
                     /label=T192A
     misc_feature    5836..5838
                     /label=K194I
     misc_feature    5881..5883
                     /label=M209V
     misc_structure  5977..5979
                     /label=M241T
     misc_feature    6043..6045
                     /label=S263P
     misc_structure  6127..6129
                     /label=G291G
     misc_feature    6169..6171
                     /label=I305V
     misc_feature    6220..6225
                     /label=NheI
     misc_feature    6226..6267
                     /label=V5 tag
     misc_feature    6274
                     /label=Geneious name: CHIMERA Pcr product - Fragment 2 ex
                     /note="Geneious type: Concatenated sequence"
     terminator      6280..6527
                     /label=CYC1_terminator
                     /note="CYC1_terminator;26to274"
     primer_bind     complement(6545..6565)
                     /label=T7
     primer_bind     complement(6574..6591)
                     /label=M13-fwd
     primer_bind     complement(6574..6590)
                     /label=M13_forward20_primer
     misc_feature    6576..6719
                     /label=lacZ_a
     primer_bind     complement(6583..6605)
                     /label=M13_pUC_fwd_primer
     CDS             6662..6730
                     /label=LacZ alpha
     rep_origin      6731..7186
                     /label=M13 origin
     rep_origin      6748..7054
                     /label=f1_origin
     rep_origin      6748..7054
                     /label=F1 ori
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 tgaaagacaa tactgtgcct
     5281 ctgaagctga tcgctctcct ggctaatggc gagttccata gtggcgaaca gctgggagaa
     5341 accctgggca tgtccagggc cgctatcaac aagcacattc agactctgcg cgactggggc
     5401 gtggacgtgt tcaccgtgcc cggaaagggc tactctctgc ccgagcctat cccgctgctg
     5461 aacgctaaac agattctggg acagctggac ggcgggagcg tggcagtcct gcctgtggtc
     5521 gactccacca atcagtacct gctggatcga atcggcgagc tgaagagtgg ggatgcttgc
     5581 attgcagaat atcagcaggc agggagagga agcagaggga ggaaatggtt ctctcctttt
     5641 ggagctaacc tgtacctgag tatgttttgg cgcctgaagc ggggaccagc agcaatcggc
     5701 ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc gaaagctggg agcagacaag
     5761 gtgcgagtca aatggcccaa tgacctgtat ctgcaggata gaaagctggc aggcatcctg
     5821 gtggagctgg ccggaataac aggcgatgct gcacagatcg tcattggcgc cgggattaac
     5881 gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa
     5941 gcagggatta acctggacag gaatactctg gccgctacgc tgatccgaga gctgcgggca
     6001 gccctggaac tgttcgagca ggaaggcctg gctccatatc tgccacggtg ggagaagctg
     6061 gataacttca tcaatagacc cgtgaagctg atcattgggg acaaagagat tttcgggatt
     6121 agccggggga ttgataaaca gggagccctg ctgctggaac aggacggagt tatcaaaccc
     6181 tggatgggcg gagaaatcag tctgcggtct gccgaaaagg ctagcggcaa gcccatcccc
     6241 aaccccctgc tgggcctgga cagcacctaa tagctcgagt catgtaatta gttatgtcac
     6301 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     6361 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     6421 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     6481 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa
     6541 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6601 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6661 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6721 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6781 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7021 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7081 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7141 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcctg atgcggtatt
     7201 ttctccttac gcatctgtgc ggtatttcac accgcatatc gacggtcgag gagaacttct
     7261 agtatatcca catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
     7321 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     7381 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     7441 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     7501 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     7561 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     7621 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     7681 ggaaaggtga gaggccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
     7741 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
     7801 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
     7861 atatatatat ttcaaggata taccattcta
//
