LOCUS       Exported                7161 bp ds-DNA     circular SYN 31-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7161)
  AUTHORS   Hampton lab
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7161
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        complement(569..587)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(608..624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    632..648
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(656..686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(1010..1598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1769..2629)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2630..2734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2766..2786
                     /label=oRH4891
     CDS             complement(2820..3623)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3624..3839)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3842..3861)
                     /label=oRH4890
     promoter        4109..4484
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4485..6200
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(6405..6860)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7005..7021
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7024..7050
                     /label=Recomb OH 1
     promoter        7031..7049
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7075..7091
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7125..7141)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7141..7161,1..9))
                     /label=Recomb OH 2
ORIGIN
        1 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
       61 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      121 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      181 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      241 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      301 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      361 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      421 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      481 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcgcggccgc
      541 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      661 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
      721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2761 acgtctgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     2821 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     2881 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     2941 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     3001 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     3061 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     3121 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc
     3181 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata
     3241 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     3301 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     3361 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa
     3421 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca
     3481 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc
     3541 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt
     3601 tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc
     3661 agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat
     3721 accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca
     3781 aaaaaatttc aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat
     3841 tgaaaagctg tggtatggtg catcatgaca ttaacctata aaaataggcg tatcacgagg
     3901 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3961 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4021 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4081 ctgagagtgc accatagatc tgaattaatt cttgaataat acataacttt tcttaaaaga
     4141 atcaaagaca gataaaattt aagagatatt aaatattagt gagaagccga gaattttgta
     4201 acaccaacat aacactgaca tctttaacaa cttttaatta tgatacattt cttacgtcat
     4261 gattgattat tacagctatg ctgacaaatg actcttgttg catggctacg aaccgggtaa
     4321 tactaagtga ttgactcttg ctgacctttt attaagaact aaatggacaa tattatggag
     4381 catttcatgt ataaattggt gcgtaaaatc gttggatctc tcttctaagt acatcctact
     4441 ataacaatca agaaaaacaa gaaaatcgga caaaacaatc aagtatggat tctagaacag
     4501 ttggtatatt aggaggggga caattgggac gtatgattgt tgaggcagca aacaggctca
     4561 acattaagac ggtaatacta gatgctgaaa attctcctgc caaacaaata agcaactcca
     4621 atgaccacgt taatggctcc ttttccaatc ctcttgatat cgaaaaacta gctgaaaaat
     4681 gtgatgtgct aacgattgag attgagcatg ttgatgttcc tacactaaag aatcttcaag
     4741 taaaacatcc caaattaaaa atttaccctt ctccagaaac aatcagattg atacaagaca
     4801 aatatattca aaaagagcat ttaatcaaaa atggtatagc agttacccaa agtgttcctg
     4861 tggaacaagc cagtgagacg tccctattga atgttggaag agatttgggt tttccattcg
     4921 tcttgaagtc gaggactttg gcatacgatg gaagaggtaa cttcgttgta aagaataagg
     4981 aaatgattcc ggaagctttg gaagtactga aggatcgtcc tttgtacgcc gaaaaatggg
     5041 caccatttac taaagaatta gcagtcatga ttgtgaggtc tgttaacggt ttagtgtttt
     5101 cttacccaat tgtagagact atccacaagg acaatatttg tgacttatgt tatgcgcctg
     5161 ctagagttcc ggactccgtt caacttaagg cgaagttgtt ggcagaaaat gcaatcaaat
     5221 cttttcccgg ttgtggtata tttggtgtgg aaatgttcta tttagaaaca ggggaattgc
     5281 ttattaacga aattgcccca aggcctcaca actctggaca ttataccatt gatgcttgcg
     5341 tcacttctca atttgaagct catttgagat caatattgga tttgccaatg ccaaagaatt
     5401 tcacatcttt ctccaccatt acaacgaacg ccattatgct aaatgttctt ggagacaaac
     5461 atacaaaaga taaagagcta gaaacttgcg aaagagcatt ggcgactcca ggttcctcag
     5521 tgtacttata tggaaaagag tctagaccta acagaaaagt aggtcacata aatattattg
     5581 cctccagtat ggcggaatgt gaacaaaggc tgaactacat tacaggtaga actgatattc
     5641 caatcaaaat ctctgtcgct caaaagttgg acttggaagc aatggtcaaa ccattggttg
     5701 gaatcatcat gggatcagac tctgacttgc cggtaatgtc tgccgcatgt gcggttttaa
     5761 aagattttgg cgttccattt gaagtgacaa tagtctctgc tcatagaact ccacatagga
     5821 tgtcagcata tgctatttcc gcaagcaagc gtggaattaa aacaattatc gctggagctg
     5881 gtggggctgc tcacttgcca ggtatggtgg ctgcaatgac accacttcct gtcatcggtg
     5941 tgcccgtaaa aggttcttgt ctagatggag tagattcttt acattcaatt gtgcaaatgc
     6001 ctagaggtgt tccagtagct accgtcgcta ttaataatag tacgaacgct gcgctgttgg
     6061 ctgtcagact gcttggcgct tatgattcaa gttatacaac gaaaatggaa cagtttttat
     6121 taaagcaaga agaagaagtt cttgtcaaag cacaaaagtt agaaactgtc ggttacgaag
     6181 cttatctaga aaacaagtaa tatataagtt tattgatata cttgtacagc aaataattat
     6241 aaaatgatat acctattttt taggctttgt tatgattaca tcaaatgtgg acttcataca
     6301 tagaaatcaa cgcttacagg tgtccttttt taagaatttc atacataaga tctatgcggt
     6361 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taaacgttaa
     6421 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6481 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6541 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6601 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6661 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6721 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6781 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6841 tgcgccgcta cagggcgcgt cgcgccattc gccattcagg ctgcgcaact gttgggaagg
     6901 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6961 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     7021 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
     7081 cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag
     7141 agcggccgcc ctcttagcgc t
//
