LOCUS       pPink-alpha-HC.        7898 bp DNA     circular SYN 13-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pPink-alpha-HC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7898)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7898)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             941..1195
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
     terminator      1248..1497
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        1517..1529
                     /gene="Pichia pastoris ADE2"
                     /label=Pichia pastoris ADE2 promoter
                     /note="PpADE2 HC promoter"
                     /note="weak 13-bp promoter fragment that requires high-copy
                     integration for efficient expression"
     CDS             1530..3218
                     /label=PpADE2
                     /note="phosphoribosylaminoimidazole carboxylase from Pichia
                     pastoris"
     promoter        3448..3597
                     /label=PpTRP2 promoter
                     /note="promoter for Pichia pastoris TRP2"
     CDS             3598..5211
                     /label=PpTRP2
                     /note="anthranilate synthase from Pichia pastoris"
     primer_bind     complement(5276..5292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5300..5316)
                     /label=lac operator
                     /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(5324..5354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5369..5390)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5678..6266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6440..7297)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7298..7402)
                     /label=AmpR promoter
     primer_bind     7876..7892
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
      961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
     1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
     1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
     1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaaggcctca
     1201 ggtaccggcc ggccatttaa atacaggccc cttttccttt gtcgatatca tgtaattagt
     1261 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     1321 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     1381 acgttattta tatttcaaat ttttcttttt tttctgtaca aacgcgtgta cgcatgtaac
     1441 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     1501 ctggatccgc ggccgccttc caaactctca tggattctca ggtaataggt attctaggag
     1561 gaggccagct aggccgaatg attgttgagg ccgctagcag gctcaatatc aagaccgtga
     1621 ttcttgatga tggtttttca cctgctaagc acattaatgc tgcgcaagac cacatcgacg
     1681 gatcattcaa agatgaggag gctatcgcca agttagctgc caaatgtgat gttctcactg
     1741 tagagattga gcatgtcaac acagatgctc taaagagagt tcaagacaga actggaatca
     1801 agatatatcc tttaccagag acaatcgaac taatcaagga taagtacttg caaaaggaac
     1861 atttgatcaa gcacaacatt tcggtgacaa agtctcaggg tatagaatct aatgaaaagg
     1921 cgctgctttt gtttggagaa gagaatggat ttccatatct gttgaagtcc cggactatgg
     1981 cttatgatgg aagaggcaat tttgtagtgg agtctaaaga ggacatcagt aaggcattag
     2041 agttcttgaa agatcgtcca ttgtatgccg agaagtttgc tccttttgtt aaagaattag
     2101 cggtaatggt tgtgagatca ctggaaggcg aagtattctc ctacccaacc gtagaaactg
     2161 tgcacaagga caatatctgt catattgtgt atgctccggc cagagttaat gacaccatcc
     2221 aaaagaaagc tcaaatatta gctgaaaaca ctgtgaagac tttcccaggc gctggaatct
     2281 tcggagttga gatgttccta ttgtctgatg gagaacttct tgtaaatgag attgctccaa
     2341 ggccccacaa ttctggtcac tatacaatcg atgcatgtgt aacatctcag ttcgaagcac
     2401 atgtaagagc cataactggt ctgccaatgc cactagattt caccaaacta tctacttcca
     2461 acaccaacgc tattatgctc aatgttttgg gtgctgaaaa atctcacggg gaattagagt
     2521 tttgtagaag agccttagaa acacccggtg cttctgtata tctgtacgga aagaccaccc
     2581 gattggctcg taagatgggt catatcaaca taataggatc ttccatgttg gaagcagaac
     2641 aaaagttaga gtacattcta gaagaatcaa cccacttacc atccagtact gtatcagctg
     2701 acactaaacc gttggttgga gttatcatgg gttcagactc tgatctacct gtgatttcga
     2761 aaggttgcga tattttaaaa cagtttggtg ttccattcga agttactatt gtctctgctc
     2821 atagaacacc acagagaatg accagatatg cctttgaagc cgctagtaga ggtatcaagg
     2881 ctatcattgc aggtgctggt ggtgctgctc atcttccagg aatggttgct gccatgactc
     2941 cgttgccagt cattggtgtt cctgtcaagg gctctacgtt ggatggtgta gactcgctac
     3001 actcgattgt ccaaatgcct agaggtgttc ctgtggctac ggttgctatc aacaacgcca
     3061 ccaatgccgc tctgttggcc atcaggattt taggtacaat tgaccacaaa tggcaaaagg
     3121 aaatgtccaa gtatatgaat gcaatggaga ccgaagtgtt ggggaaggca tccaacttgg
     3181 aatctgaagg gtatgaatcc tatttgaaga atcgtctttg aatttagtat tgttttttaa
     3241 tagatgtata tataatagta cacgtaactt atctattcca ttcataattt tattttaaag
     3301 gttcggtaga aatttgtcct ccaaaaagtt ggttagagcc tggcagtttt gataggcatt
     3361 attatagatt gggtaatatt taccctgcac ctggaggaac tttgcaaaga gcctcatgtg
     3421 cggcgcgcca ggccataatg gccaaacggt ttctcaatta ctatatacta ctaaccattt
     3481 acctgtagcg tatttctttt ccctcttcgc gaaagctcaa gggcatcttc ttgactcatg
     3541 aaaaatatct ggatttcttc tgacagatca tcacccttga gcccaactct ctagcctatg
     3601 agtgtaagtg atagtcatct tgcaacagat tattttggaa cgcaactaac aaagcagata
     3661 cacccttcag cagaatcctt tctggatatt gtgaagaatg atcgccaaag tcacagtcct
     3721 gagacagttc ctaatcttta ccccatttac aagttcatcc aatcagactt cttaacgcct
     3781 catctggctt atatcaagct taccaacagt tcagaaactc ccagtccaag tttcttgctt
     3841 gaaagtgcga agaatggtga caccgttgac aggtacacct ttatgggaca ttcccccaga
     3901 aaaataatca agactgggcc tttagagggt gctgaagttg accccttggt gcttctggaa
     3961 aaagaactga agggcaccag acaagcgcaa cttcctggta ttcctcgtct aagtggtggt
     4021 gccataggat acatctcgta cgattgtatt aagtactttg aaccaaaaac tgaaagaaaa
     4081 ctgaaagatg ttttgcaact tccggaagca gctttgatgt tgttcgacac gatcgtggct
     4141 tttgacaatg tttatcaaag attccaggta attggaaacg tttctctatc cgttgatgac
     4201 tcggacgaag ctattcttga gaaatattat aagacaagag aagaagtgga aaagatcagt
     4261 aaagtggtat ttgacaataa aactgttccc tactatgaac agaaagatat tattcaaggc
     4321 caaacgttca cctctaatat tggtcaggaa gggtatgaaa accatgttcg caagctgaaa
     4381 gaacatattc tgaaaggaga catcttccaa gctgttccct ctcaaagggt agccaggccg
     4441 acctcattgc accctttcaa catctatcgt catttgagaa ctgtcaatcc ttctccatac
     4501 atgttctata ttgactatct agacttccaa gttgttggtg cttcacctga attactagtt
     4561 aaatccgaca acaacaacaa aatcatcaca catcctattg ctggaactct tcccagaggt
     4621 aaaactatcg aagaggacga caattatgct aagcaattga agtcgtcttt gaaagacagg
     4681 gccgagcacg tcatgctggt agatttggcc agaaatgata ttaaccgtgt gtgtgagccc
     4741 accagtacca cggttgatcg tttattgact gtggagagat tttctcatgt gatgcatctt
     4801 gtgtcagaag tcagtggaac attgagacca aacaagactc gcttcgatgc tttcagatcc
     4861 attttcccag caggaaccgt ctccggtgct ccgaaggtaa gagcaatgca actcatagga
     4921 gaattggaag gagaaaagag aggtgtttat gcgggggccg taggacactg gtcgtacgat
     4981 ggaaaatcga tggacacatg tattgcctta agaacaatgg tcgtcaagga cggtgtcgct
     5041 taccttcaag ccggaggtgg aattgtctac gattctgacc cctatgacga gtacatcgaa
     5101 accatgaaca aaatgagatc caacaataac accatcttgg aggctgagaa aatctggacc
     5161 gataggttgg ccagagacga gaatcaaagt gaatccgaag aaaacgatca atgaacggag
     5221 gacgtaagta ggaatttatg gtttggccat aatggcctag cttggcgtaa tcatggtcat
     5281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6541 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7261 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7321 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7381 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7441 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7501 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     7561 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7621 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7681 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     7741 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7801 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7861 cccagtcacg acgttgtaaa acgacggcca gtgaattg
//
