LOCUS       pBS-SKII-URA-GAL        5454 bp DNA     circular SYN 16-FEB-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5454)
  AUTHORS   NP
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5454
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     complement(1..17)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(54..72)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(89..107)
                     /label=O#1477
     primer_bind     complement(93..109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(107..128)
                     /label=O#4002
     protein_bind    117..133
                     /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(141..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    186..207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(495..1083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1254..2114)
                     /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(2115..2219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2245..2700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2842..2858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2868..2887
                     /label=O#40
     promoter        2868..2886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2912..2928
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2948..4346
                     /label=C. albicans URA3
     promoter        2963..3306
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4125..4322
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4327..4366)
                     /label=GAL1pro-TEFt-rev
     promoter        4347..4788
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4347..4464
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4765..4788)
                     /label=GAL1pro-rev
     primer_bind     4769..4808
                     /label=GAL1pro-SNAP/CLIP-fwd
     CDS             4789..5334
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(4875..4898)
                     /label=O#2951
     primer_bind     5315..5354
                     /label=SNAP/CLIP-3xHA-fwd
     primer_bind     complement(5318..5334)
                     /label=SNAP/CLIP-rev
     CDS             5335..5424
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5434..5454
                     /label=GSGGSGS linker
     primer_bind     5434..5453
                     /label=O#4211
ORIGIN
        1 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
       61 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
      121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
      961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2221 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     2281 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2341 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2401 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2461 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2521 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2581 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2641 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2701 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2761 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2821 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     2881 tatagggcga attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata
     2941 tcgaattggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     3001 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     3061 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     3121 ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     3181 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     3241 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     3301 acaaccatga cagtcaacac taagacctat agtgagagag cagaaactca tgcctcacca
     3361 gtagcacaac gattatttcg attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt
     3421 gatgttgata ccactaagga attccttgaa ttaattgata aattgggtcc ttatgtatgc
     3481 ttaatcaaga ctcatattga tataatcaat gatttttcct atgaatccac tattgaacca
     3541 ttattagaac tttcacgtaa acatcaattt atgatttttg aagatagaaa atttgctgat
     3601 attggtaata ccgtgaagaa acaatatatt ggtggagttt ataaaattag tagttgggca
     3661 gatattacta atgctcatgg tgtcactggg aatggagtag ttgaaggatt aaaacaggga
     3721 gctaaagaaa ccaccaccaa ccaagagcca agagggttat tgatgttagc tgaattatca
     3781 tcagtgggat cattagcata tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc
     3841 gataaggaat ttgttattgg atttattgcc caacgtgata tgggtggaca agaagaagga
     3901 tttgattggc ttattatgac acctggagtt ggattagatg ataaaggtga tggattagga
     3961 caacaatata gaactgttga tgaagttgtt agcactggaa ctgatattat cattgttggt
     4021 agaggattgt ttggtaaagg aagagatcca gatattgaag gtaaaaggta tagagatgct
     4081 ggttggaatg cttatttgaa aaagactggc caattataat cagtactgac aataaaaaga
     4141 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     4201 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     4261 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     4321 tgctgtcgat tcgatactaa cgccgccgga ttagaagccg ccgagcgggt gacagccctc
     4381 cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga
     4441 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
     4501 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat
     4561 taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc
     4621 agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact gcataaccac
     4681 tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat gtaataaaag
     4741 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagat ggacaaagac
     4801 tgcgaaatga agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc
     4861 gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg
     4921 gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc cactgatcca ggccaccgcc
     4981 tggctcaacg cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg
     5041 caccacccag tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa
     5101 gtggtgaagt tcggagaggt catcagcgag agccacctgg ccgccctggt gggcaatccc
     5161 gccgccaccg ccgccgtgaa caccgccctg gacggaaatc ccgtgcccat tctgatcccc
     5221 tgccaccggg tggtgcaggg cgacagcgac gtggggccct acctgggcgg gctcgccgtg
     5281 aaagagtggc tgctggccca cgagggccac agactgggca agcctgggct gggttaccca
     5341 tacgatgttc ctgactatgc gggctatccc tatgacgtcc cggactatgc gggatcctat
     5401 ccatatgacg ttccagatta cgctgctcag tgcggttcag gtggatctgg ttct
//
