LOCUS       Exported                5286 bp ds-DNA     circular SYN 28-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5286)
  AUTHORS   Macapp
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 28, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5286
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        1..528
                     /label=AOX_RIGHT
     CDS             535..1527
                     /codon_start=1
                     /label=mGIRK2
                     /translation="MAKRKIQRYVRKDGKCNVHHGNVRETYRYLTDIFTTLVDLKWRFN
                     LLIFVMVYTVTWLFFGMIWWLIAYIRGDMDHIEDPSWTPCVTNLNGFVSAFLFSIETET
                     TIGYGYRVITDKCPEGIILLLIQSVLGSIVNAFMVGCMFVKISQPKKRAETLVFSTHAV
                     ISMRDGKLCLMFRVGDLRNSHIVEASIRAKLIKSKQTSEGEFIPLNQSDINVGYYTGDD
                     RLFLVSPLIISHEINQQSPFWEISKAQLPKEELEIVVILEGIVEATGMTCQARSSYITS
                     EILWGYRFTPVLTMEDGFYEVDYNSFHETYETSTPSLSAKELAELANRAE"
     CDS             1537..1557
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1564..2268
                     /codon_start=1
                     /label=mCherry_pichia_codon_optimized
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     CDS             2284..2307
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      2387..2633
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2648..3059
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3067..3114
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3133..3507
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3573..3820
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    3877..3953
                     /label=BxB1 attP
     misc_feature    3993..4056
                     /label=R4 attP
     misc_feature    4107..4175
                     /label=TP901 attP
     promoter        4230..4645
                     /label=AOX_Left
     rep_origin      complement(4673..5261)
                     /direction=LEFT
                     /label=ori_pPIC
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtttaaacgc tgtcttggaa cctaatatga caaaagcgtg atctcatcca agatgaacta
       61 agtttggttc gttgaaatgc taacggccag ttggtcaaaa agaaacttcc aaaagtcggc
      121 ataccgtttg tcttgtttgg tattgattga cgaatgctca aaaataatct cattaatgct
      181 tagcgcagtc tctctatcgc ttctgaaccc cggtgcacct gtgccgaaac gcaaatgggg
      241 aaacacccgc tttttggatg attatgcatt gtctccacat tgtatgcttc caagattctg
      301 gtgggaatac tgctgatagc ctaacgttca tgatcaaaat ttaactgttc taacccctac
      361 ttgacagcaa tatataaaca gaaggaagct gccctgtctt aaaccttttt ttttatcatc
      421 attattagct tactttcata attgcgactg gttccaattg acaagctttt gattttaacg
      481 acttttaacg acaacttgag aagatcaaaa aacaactaat tattcgaaac gaaaatggca
      541 aaacgtaaga tccaacgtta cgttcgtaag gacggtaagt gcaatgtgca tcatggtaat
      601 gtaagagaaa cctacagata cctaacggac attttcacca ctttggtgga tttgaaatgg
      661 cgtttcaact tattgatatt tgttatggta tacaccgtta cgtggctatt tttcggtatg
      721 atttggtggc taattgcata tataagaggt gacatggatc acatcgaaga cccctcctgg
      781 actccatgtg ttaccaatct aaacggcttt gtttccgcat tcttgttttc catcgaaact
      841 gagaccacaa ttggttacgg ttaccgtgta attacagata aatgcccaga gggtatcatc
      901 ttgttactaa ttcaatccgt tttaggctcc attgttaatg ctttcatggt cggttgcatg
      961 tttgtgaaaa tttctcagcc caaaaagagg gctgaaactc ttgttttctc cacacatgca
     1021 gttatatcaa tgagggatgg caagttatgc ctgatgtttc gtgtcggaga cctaagaaat
     1081 tcacatatag tggaagcaag tattagagct aaactgataa aaagtaagca aacatccgag
     1141 ggcgagttca tcccattgaa tcagtctgac attaacgtgg gatactatac tggcgacgac
     1201 cgtctttttt tagttagtcc acttatcatt tcccacgaaa ttaatcagca gtcccctttt
     1261 tgggagattt ccaaggctca attgcctaag gaagaactag agattgtcgt gattcttgaa
     1321 ggcatcgtcg aagccactgg catgacgtgc caagcacgtt cctcttacat cacatccgag
     1381 atactatggg gatatagatt cacgcccgtc ttaaccatgg aagatggatt ctacgaagta
     1441 gattataaca gtttccatga gacctatgag acgagtactc cttccctatc tgcaaaggaa
     1501 ttggctgaac tggctaatag agccgagggt cctggcgaaa acctgtattt tcagggtact
     1561 agtgtctcta agggagaaga ggacaatatg gctattatca aggagttcat gagatttaaa
     1621 gtccatatgg agggctctgt taatggtcat gaatttgaaa ttgaaggtga aggagaagga
     1681 agaccttatg agggaaccca aacggctaag ctgaaagtaa ctaaaggcgg cccacttcct
     1741 ttcgcatggg atatactgtc tccacaattc atgtatggct ctaaagctta tgtaaagcac
     1801 cccgccgaca ttcccgatta tctaaagtta tcctttcccg agggatttaa atgggaaagg
     1861 gttatgaact tcgaggatgg cggcgtggta actgttacgc aggactcctc ccttcaagat
     1921 ggtgaattta tttacaaagt gaagcttcgt ggtactaatt tcccaagtga cggtcctgtg
     1981 atgcagaaga agactatggg ttgggaagct agttccgaga ggatgtaccc cgaagatggt
     2041 gccctgaagg gtgagattaa gcaacgtcta aaattaaaag acggcggtca ctatgacgct
     2101 gaagtaaaga ccacatataa ggcaaaaaag cccgtccagc ttcctggcgc ctacaatgtt
     2161 aatatcaagt tggacatcac ctcccataac gaagattaca ctatcgtcga gcaatacgag
     2221 agggcagagg gaagacattc cacaggtggt atggacgagc tatataaact cgagggcgct
     2281 tcctggtcac atccacagtt tgagaaatga gtttgtagcc ttagacatga ctgttcctca
     2341 gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca
     2401 gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta
     2461 tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag
     2521 cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga
     2581 tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt
     2641 ttgtgcggat cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc
     2701 agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact
     2761 aaattttccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa
     2821 agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat
     2881 cacgtttctt tttcttgaaa tttttttttt tagttttttt ctctttcagt gacctccatt
     2941 gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc
     3001 tattacaact ttttttactt cttgttcatt agaaagaaag catagcaatc taatctaagg
     3061 ggcggtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3121 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     3181 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     3241 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     3301 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3361 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     3421 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3481 cacttcgtgg ccgaggagca ggactgacac gtccgacggc ggcccacggg tcccaggcct
     3541 cggagatccg tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
     3601 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     3661 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     3721 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     3781 gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg
     3841 agcaaaaggc cagggcgacg taaacggcca caagtttatg gccgtgatga cctgtgtctt
     3901 cgtggtttgt ctggtcaacc accgcggtct cagtggtgta cggtacaaac ccaaagcagc
     3961 acgacacggc aactacaaga cccgcgccga gggcatgttc cccaaagcga taccacttga
     4021 agcagtggta ctgcttgtgg gtacactctg cgggtgtgaa gttcgagggc gacaccctgg
     4081 tgaaccgcat cgagctgaag ggcatcttcc aactcgctta attgcgagtt tttatttcgt
     4141 ttatttcaat taaggtaact aaaaaactcc ttttacacat gaagcagcac gacttcttca
     4201 agcgttaagg gattttggtc atgagatcag atctaacatc caaagacgaa aggttgaatg
     4261 aaaccttttt gccatccgac atccacaggt ccattctcac acataagtgc caaacgcaac
     4321 aggaggggat acactagcag cagaccgttg caaacgcagg acctccactc ctcttctcct
     4381 caacacccac ttttgccatc gaaaaaccag cccagttatt gggcttgatt ggagctcgct
     4441 cattccaatt ccttctatta ggctactaac accatgactt tattagcctg tctatcctgg
     4501 cccccctggc gaggttcatg tttgtttatt tccgaatgca acaagctccg cattacaccc
     4561 gaacatcact ccagatgagg gctttctgag tgtggggtca aatagtttca tgttccccaa
     4621 atggcccaaa actgacagtt taaacccgta aaaaggccgc gttgctggcg tttttccata
     4681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4861 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5101 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5281 ctacgg
//
