LOCUS       Exported                5593 bp ds-DNA     circular SYN 10-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5593)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 11, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5593
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             216..249
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAC"
     misc_feature    233..249
                     /label=MCS
                     /note="pUC19 multiple cloning site"
     primer_bind     244..270
                     /label=MUP1.FOR-SbfI
     CDS             250..870
                     /codon_start=1
                     /label=MUP1
                     /translation="GREGVLPFSNFFASSKPFNSPMVGLFQHFIVCTVTILAPPPGDAY
                     LLVQNLISYPMNIINFAISAGLLWIYWQRRQGKIEWNPPIKAGVFVTGFFTLSNLYLII
                     APYVPPSNGESVYSSMPYWIHCVIAWGIFFFGGVYYVVWAQLLPRWGHYKLVSKDVLGE
                     DGFWRVKIAKVYDDTIGDVDTQEDGVIETNIIEHYKSEQEKSLK"
     primer_bind     complement(850..874)
                     /label=MUP1.BGLIIN.REV
     primer_bind     871..891
                     /label=yo-mScarlet2i final in-frame QC2.FOR
     misc_feature    871..874
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     CDS             883..1572
                     /codon_start=1
                     /label=yo-mScarlet2-I
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGSGAS"
     primer_bind     1572..1602
                     /label=MUP1.BGLIIN.FOR
     misc_feature    1581..2076
                     /label=3' MUP1
     primer_bind     complement(2050..2079)
                     /label=MUP1.REV-SacI
     primer_bind     2075..2096
                     /label=BV13
     CDS             2079..2335
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="NSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQ
                     QLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     misc_feature    2079..2085
                     /label=MCS
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(2086..2102)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2105..2126
                     /label=YIplac211.SEQ.1 BU28
     CDS             complement(2570..3373)
                     /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"
     primer_bind     3042..3066
                     /label=YIplac211.SEQ.2 BU29
     promoter        complement(3374..3594)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3792..4652
                     /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"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gacgtgaagg tgtgttgccc ttttctaatt tctttgcctc ttctaagcca
      301 ttcaactccc caatggttgg tttgtttcaa cattttattg tttgtacagt aaccatttta
      361 gccccacctc caggtgatgc atatttgcta gtccaaaatt taatttccta cccaatgaat
      421 attattaatt ttgccattag cgcgggtttg ctttggatat actggcaacg tagacaaggt
      481 aaaattgaat ggaacccacc aattaaggcc ggtgtttttg ttacgggatt tttcacatta
      541 tccaatttat acctgattat tgctccttac gttccacctt caaatggcga gtcggtgtat
      601 tcaagcatgc catactggat ccactgtgta attgcttggg gtattttctt ctttggtggt
      661 gtttactacg ttgtttgggc ccaattattg ccaagatggg gccactacaa attggtttcc
      721 aaggatgtgc ttggtgaaga tgggttctgg agagtcaaaa ttgccaaagt ttatgatgac
      781 actattggcg atgtcgatac acaagaagac ggcgttattg aaacgaatat aatcgaacat
      841 tacaaaagtg aacaagaaaa atcgctgaaa gatccaacta gtatggatag tacagaagca
      901 gttataaagg aattcatgag attcaaagtc cacatggaag gtagtatgaa cggtcacgaa
      961 ttcgaaatag aaggtgaagg tgaaggtaga ccatatgaag gtactcaaac agctaaattg
     1021 aaggttacta aaggtggtcc attgccattt tcttgggata tcttgtctcc acaattcatg
     1081 tacggttcaa gagcttttat taaacatcca gcagatattc cagattacta caagcaatca
     1141 tttccagaag gttttaaatg ggaaagagtt atgaacttcg aagatggtgg tgcagttact
     1201 gttacacaag atacttcttt ggaagatggt acattgatct ataaggttaa attgagaggt
     1261 acaaattttc caccagatgg tccagttatg caaaagaaaa ctatgggttg ggaagcatct
     1321 acagaaagat tgtacccaga agatggtgtt ttgaagggtg acatcaagat ggcattgaga
     1381 ttgaaggatg gtggtagata tttggctgat ttcaagacta catacaaggc taagaaacca
     1441 gttcaaatgc caggtgctta caacgttgat agaaaattgg atatcacttc tcataacgaa
     1501 gattacacag ttgttgaaca atacgaaaga agtgaaggta gacatagtac aggtggtagt
     1561 ggtgctagct aagcggccgc ccaaaaaggt acttaccatc tcatctcgat tcttataatc
     1621 acgtatgaac attcttataa ttgcattttc tttatacaaa atatataggt accaaacctc
     1681 attataattt ctgccattta atagtttaag atgctattcc gtttcgtatt ttttcaattt
     1741 ttatacctac tattgcgcga aatcaaaata gcatactaag aaaaagcaat atcatgaatt
     1801 gctgatgaga tagattcaaa ttggaattaa ctaaatctat cagcaacgct ataaacacag
     1861 ataagcgctc tataaaatcg tttgtctaac gttttatata gttctttatt gctcccttac
     1921 cctctttccg tcaatagtga ttgttgtctc gggaaagctt ttttttttat taatcaaagg
     1981 cccaattgag ttacaactga aattaagctg agttggcgga acagtgtggt attagcgaag
     2041 ggaaaatctg tgaagtgccc tagctacaaa acagagctcg aattcactgg ccgtcgtttt
     2101 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2161 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2221 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2281 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2341 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2401 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2461 gtcatcaccg aaacgcgcga gacgaaaggg cgggtaataa ctgatataat taaattgaag
     2521 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     2581 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     2641 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     2701 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     2761 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     2821 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     2881 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     2941 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     3001 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     3061 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     3121 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     3181 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     3241 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     3301 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     3361 tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta
     3421 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     3481 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     3541 aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg
     3601 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga aga
//
