LOCUS       Exported                6894 bp DNA     circular SYN 20-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2022 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6894
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(50..69)
                     /label=Rseq_ScURA3t
     primer_bind     complement(235..251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    259..275
                     /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(283..313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    328..349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      637..1225
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1396..2256)
                     /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(2257..2361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2835..2851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2855..3027
                     /label=S. cerevisiae URA3 promoter
     gene            3028..3831
                     /product="Kluyveromyces lactis URA3"
                     /label=K. lactis URA3
     terminator      3832..3949
                     /label=K. lactis URA3 terminator
     primer_bind     3884..3903
                     /label=Rseq_KlURA3t
     primer_bind     3935..3987
                     /label=HIS5t_KlURA3t_rv
     terminator      3961..4105
                     /label=HIS5 terminator
     primer_bind     4076..4123
                     /label=APEX2_HIS5t_rv
     CDS             complement(4106..4858)
                     /codon_start=1
                     /product="Engineered ascorbate peroxidase (APEX2) from ref.
                     Lam et al. 2015. Used as an in vivo TEM reporter. "
                     /label=APEX2
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     primer_bind     4108..4124
                     /label=APEX_mODC_rv_long
     primer_bind     complement(4837..4871)
                     /label=APEX2_VP2_fw
     gene            complement(4859..5056)
                     /label=VP2C
     primer_bind     complement(5036..5095)
                     /label=VP2_ClaI_GAL10p_fw
     promoter        5066..5733
                     /label=GAL10/GAL1 bi-directional promoter
                     /note="Obtained from plasmid pILGFPB5A"
     primer_bind     complement(5118..5148)
                     /label=Fseq_GAL10p
     protein_bind    5281..5398
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5650..5681
                     /label=Fseq_GAL1p
     primer_bind     5707..5759
                     /label=VP1_NheI_GAL1p_fw
     gene            5743..6885
                     /label=deltaVP1
     primer_bind     complement(join(6860..6894,1..21))
                     /label=VP1_NotI_URA3t_rv
     terminator      join(6894,1..234)
                     /label=S. cerevisiae URA3 terminator
ORIGIN
        1 ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
       61 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
      121 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
      181 cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat catggtcata
      241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     2461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2821 ccagtcacga cgttgtaaaa cgacggccag tgaattcatt taaatttttt tgattcggta
     2881 atctccgagc agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2941 atatgtggtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     3001 aaaacctgca ggaaacgaag ataaaacatg tccacaaaat catataccag tagagctgag
     3061 actcatgcaa gtccggttgc atcgaaactt ttacgtttaa tggatgaaaa gaagaccaat
     3121 ttgtgtgctt ctcttgacgt tcgttcgact gatgagctat tgaaacttgt tgaaacgttg
     3181 ggtccataca tttgcctttt gaaaacacac gttgatatct tggatgattt cagttatgag
     3241 ggtactgtcg ttccattgaa agcattggca gagaaataca agttcttgat atttgaggac
     3301 agaaaattcg ccgatatcgg taacacagtc aaattacaat atacatcggg cgtttaccgt
     3361 atcgcagaat ggtctgatat caccaacgcc cacggggtta ctggtgctgg tattgttgct
     3421 ggcttgaaac aaggtgcgca agaggtcacc aaagaaccaa ggggattatt gatgcttgct
     3481 gaattgtctt ccaagggttc tctagcacac ggtgaatata ctaagggtac cgttgatatt
     3541 gcaaagagtg ataaagattt cgttattggg ttcattgctc agaacgatat gggaggaaga
     3601 gaagaagggt ttgattggct aatcatgacc ccaggtgtag gtttagacga caaaggcgat
     3661 gcattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc agatatcatc
     3721 attgttggca gaggactttt cgccaagggt agagatccta aggttgaagg tgaaagatac
     3781 agaaatgctg gatgggaagc gtaccaaaag agaatcagcg ctccccatta attatacagg
     3841 aaacttaata gaacaaatca catatttaat ctaatagcca cctgcattgg cacggtgcaa
     3901 cactcacttc aacttcatct tacaaaagat cacgtgatct gttgtattgg gatctctaga
     3961 gtaacaatat catgagacct tttatagaag tggcgccaaa actaaatgta tttgaaaata
     4021 caaaaaacgc acaggtttct ttatatatgt atatatatat atatatatat ataaaactgt
     4081 acatatactg tttaaattaa tctatttaag catctgcaaa tcctaattct gacaatttct
     4141 ggtgggcctc tgcataatcg gcaaaaaatg cgtcctcatc agctgcatat ttatcaacaa
     4201 gtggacgaaa gacgggatca gataataaag ctttatcgga tggtaattgt aacaaacctt
     4261 ctttttcacc agataacaac tcagtaaaat aagagttatc aaatatcaaa ggattagatg
     4321 tccatggtcc ttcgaatcct gatctttctt tgtgtgcagc tccaattgta tgaccaccag
     4381 acaatgcaac tatatcttgg tctgtcaatc ccatagcttt accgaaaaca tctcttagat
     4441 gatcagagcc ttttgttgga tctggcaatc taccttcagg tggaggttct ggtttatctt
     4501 ctctacctgg atggaatgga acttttggtc ctccagtgac ttcgacagca acaacaccag
     4561 ccaattgata aaaatcagca taagacaaaa ttggaaactc agcttttaat ggttctaata
     4621 atctaactgc tatatccaaa ccattatttg cagaatgagc taattcagct ggatgtttaa
     4681 tagtaccaaa tggaccacct gtttttgtac ctttatcaaa agtaccagct gaatgaaaag
     4741 ctaatctcaa cattaatgga gcgcatcttt tttcagcaat gaaacctctc aacttctttt
     4801 ttgctttttc gacagcgtct tgatagtctg ctgaaacagt agggtatgat ttacccatag
     4861 aaccgccgcc accggagcca ccgccaccag aaccaccacc acctggagtg atgtcaccat
     4921 acaaacccaa aatcaatggc aacatccaat ctggagtaac tctttgatga gaaacaactt
     4981 gagcttgata aaatttaata acttcaccag attcatcttg aatatgagca gctggagttg
     5041 gaccaccatt acccatttta tcgatttata ttgaattttc aaaaattctt actttttttt
     5101 tggatggacg caaagaagtt taataatcat attacatggc attaccacca tatacatatc
     5161 catatctaat cttacttata tgttgtggaa atgtaaagag ccccattatc ttagcctaaa
     5221 aaaaccttct ctttggaact ttcagtaata cgcttaactg ctcattgcta tattgaagta
     5281 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
     5341 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
     5401 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
     5461 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
     5521 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
     5581 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
     5641 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
     5701 tctatacttt aacgtcaagg agaaaaaact atagctagca aaatggcttc aggtgtaagt
     5761 aaatgcgaaa ccaaatgcac aaaggcttgc cctagaccag ctccagttcc taagttattg
     5821 attaaaggtg gtatggaagt tttggatttg gttacaggtc cagattctgt tactgaaatc
     5881 gaagcatttt tgaacccaag aatgggtcaa ccaccaacac cagaatcttt aactgaaggt
     5941 ggtcaatatt acggttggtc aagaggtatt aatttggcta catctgatac tgaagattca
     6001 ccaggtaata atacattacc aacttggtct atggcaaaat tgcaattacc aatgttgaac
     6061 gaagatttga cttgtgatac tttgcaaatg tgggaagcag tttcagttaa aacagaagtt
     6121 gttggttctg gttctttgtt ggatgttcat ggttttaata agccaacaga tactgttaac
     6181 acaaagggta tttctactcc agttgaaggt tcacaatatc atgtttttgc tgttggtggt
     6241 gaaccattgg atttgcaagg tttagttaca gatgcaagaa ctaagtacaa ggaagaaggt
     6301 gttgttacta ttaaaacaat cactaagaaa gatatggtta ataaggatca agttttgaac
     6361 ccaatttcaa aggctaaatt ggataaggat ggcatgtacc cagttgaaat ttggcatcca
     6421 gatccagcta aaaatgaaaa cacaagatac ttcggtaatt acactggtgg tactacaact
     6481 ccaccagttt tgcaattcac taacactttg acaactgttt tgttagatga aaatggtgtt
     6541 ggtccattat gtaaaggtga aggtttgtac ttatcttgcg ttgatattat gggttggaga
     6601 gttacaagaa actacgatgt tcatcattgg agaggtttgc caagatactt caagattact
     6661 ttaagaaaaa gatgggttaa aaatccatac ccaatggctt ctttgatttc ttctttgttt
     6721 aataatatgt taccacaagt tcaaggtcaa ccaatggaag gtgaaaacac acaagttgaa
     6781 gaagttagag tttacgatgg tactgaacca gttccaggtg accctgacat gacaagatat
     6841 gtagatagat tcggtaaaac taaaactgta ttccctggta attaagcggc cgca
//
