LOCUS       Exported                6924 bp DNA     circular SYN 09-OCT-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  AUTHORS   Rob Mitra Lab WUSTL
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2022 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6924
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    187..1281
                     /locus_tag="LEU2 ORF"
                     /label=LEU2 ORF
     rep_origin      complement(1380..1835)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
     CDS             complement(1618..2017)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="*VVLRALTGRRFTTS*LGKPWRYPT*SPCSTSPFRQLA**RRGPH
                     RSPFPTVAQPEWRMARRAL*RRIKRGGCGGYAQRDRYTCQRPSARSFRFLPFLSRHVRR
                     LSPSSSKSGAPFRVPI*CFTAPRPQKT*L"
     primer_bind     1980..1996
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2006..2017
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2018..2037
                     /label=20bp_constant_seq
     misc_feature    2018..2029
                     /label=New Feature
     primer_bind     2018..2025
                     /label=T7
     CDS             complement(2120..2139)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="SSTAVA"
     misc_feature    2120..2139
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    2141..2157
                     /label=17bp_constant_seq
                     /note="part of the reverse primer
                     "
     misc_feature    2158..2240
                     /label=HIS3
     primer_bind     2238..2260
                     /label=Fragment 2.FOR
     CDS             2241..2957
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2937..2999)
                     /label=Fragment 2.REV
     primer_bind     2980..3028
                     /label=Fragment 3.FOR
     terminator      2981..3168
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. 
                     cerevisiaealcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3141..3168)
                     /label=Fragment 3.REV
     rep_origin      complement(3254..3842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3885..3914)
                     /label=Vector.REV
     misc_feature    3914..4467
                     /label=tef2 promoter
     misc_feature    3914
                     /label=TEF1p Promoter
     CDS             4468..5178
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein(Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4468..4487
                     /label=Fragment 2.FOR
     primer_bind     complement(5155..5197)
                     /label=Fragment 2.REV
     primer_bind     5167..5226
                     /label=Fragment 3.FOR
     terminator      5179..5366
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. 
                     cerevisiaealcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5339..5388)
                     /label=Fragment 3.REV
     primer_bind     5367..5388
                     /label=Vector.FOR
     CDS             complement(5459..6271)
                     /label=KanR
     primer_bind     complement(6317..6327)
                     /label=Vector.FOR
     misc_feature    6352..6855
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to 
                     anautonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatattaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttcggtggt
      241 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc
      301 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc
      361 agacaagata gtggcgatag ggtcaacctt attctttggc aaatctggag cagaaccgtg
      421 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga
      481 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa
      541 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc
      601 ggcagaatca atcaattgat gttgaacctt caatgtaggg aattcgttct tgatggtttc
      661 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca
      721 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg
      781 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt
      841 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt
      901 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg
      961 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct
     1021 aacactaccg gtaccccatt taggaccacc cacagcacct aacaaaacgg catcaacctt
     1081 cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga tagcagcacc
     1141 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt
     1201 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac
     1261 gatcttctta ggggcagaca tgttattact gagtagtatt tatttaagta ttgtttgtgc
     1321 acttgcctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     1381 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     1561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     1621 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1681 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1741 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1801 acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg
     1861 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     1921 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     1981 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     2041 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2101 atccactagt tctagagcgg ccgccaccgc ggtggagctc attggcatta tcacataatg
     2161 aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa aatgagcagg
     2221 caagataaac gaaggcaaag atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2281 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2341 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2401 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc
     2461 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2521 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2581 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2641 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2701 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2761 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2821 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     2881 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2941 tggacgagct gtacaagtaa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     3001 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     3061 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     3121 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggaa tcaggggata
     3181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3661 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3901 aagggatttt ggtctatact tacatatagt agatgtcaag cgtaggcgct tcccctgccg
     3961 gctgtgaggg cgccataacc aaggtatcta tagaccgcca atcagcaaac tacctccgta
     4021 cattcatgtt gcacccacac atttatacac ccagaccgcg acaaattacc cataaggttg
     4081 tttgtgacgg cgtcgtacaa gagaacgtgg gaacttttta ggctcaccaa aaaagaaaga
     4141 aaaaatacga gttgctgaca gaagcctcaa gaaaaaaaaa attcttcttc gactatgctg
     4201 gaggcagaga tgatcgagcc ggtagttaac tatatatagc taaattggtt ccatcacctt
     4261 cttttctggt gtcgctcctt ctagtgctat ttctggcttt tcctattttt ttttttccat
     4321 ttttctttct ctctttctaa tatataaatt ctcttgcatt ttctattttt ctctctatct
     4381 attctacttg tttattccct tcaaggtttt tttttaagga gtacttgttt ttagaatata
     4441 cggtcaacga actataatta actaaacatg gtgagcaagg gcgaggagga taacatggcc
     4501 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     4561 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     4621 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     4681 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     4741 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     4801 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     4861 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4921 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     4981 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     5041 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     5101 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     5161 gacgagctgt acaagtgagc gaatttctta tgatttatga tttttattat taaataagtt
     5221 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     5281 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
     5341 gctcttattg accacacctc taccggcacc tagatcccgg ggaaatgtgc gcggaacccc
     5401 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     5461 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5521 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5581 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5641 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5701 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5761 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5821 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     5881 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     5941 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     6001 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     6061 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6121 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     6181 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     6241 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6301 cagacagttt tattgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc
     6361 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     6421 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     6481 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     6541 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     6601 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     6661 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     6721 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     6781 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     6841 tacaggcaag cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata
     6901 ggcgtatcac gaggcccttt cgtc
//
