LOCUS       pPink-HC.        7667 bp DNA     circular SYN 13-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pPink-HC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7667)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     terminator      1017..1266
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        1286..1298
                     /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             1299..2987
                     /label=PpADE2
                     /note="phosphoribosylaminoimidazole carboxylase from Pichia
                     pastoris"
     promoter        3217..3366
                     /label=PpTRP2 promoter
                     /note="promoter for Pichia pastoris TRP2"
     CDS             3367..4980
                     /label=PpTRP2
                     /note="anthranilate synthase from Pichia pastoris"
     primer_bind     complement(5045..5061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5069..5085)
                     /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(5093..5123)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5138..5159)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5447..6035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6209..7066)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7067..7171)
                     /label=AmpR promoter
     primer_bind     7645..7661
                     /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 gaattccgga ccggcatgcc
      961 aaggcctcag gtaccggccg gccatttaaa tacaggcccc ttttcctttg tcgatatcat
     1021 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     1081 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     1141 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa acgcgtgtac
     1201 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     1261 atttgcaagc tggatccgcg gccgccttcc aaactctcat ggattctcag gtaataggta
     1321 ttctaggagg aggccagcta ggccgaatga ttgttgaggc cgctagcagg ctcaatatca
     1381 agaccgtgat tcttgatgat ggtttttcac ctgctaagca cattaatgct gcgcaagacc
     1441 acatcgacgg atcattcaaa gatgaggagg ctatcgccaa gttagctgcc aaatgtgatg
     1501 ttctcactgt agagattgag catgtcaaca cagatgctct aaagagagtt caagacagaa
     1561 ctggaatcaa gatatatcct ttaccagaga caatcgaact aatcaaggat aagtacttgc
     1621 aaaaggaaca tttgatcaag cacaacattt cggtgacaaa gtctcagggt atagaatcta
     1681 atgaaaaggc gctgcttttg tttggagaag agaatggatt tccatatctg ttgaagtccc
     1741 ggactatggc ttatgatgga agaggcaatt ttgtagtgga gtctaaagag gacatcagta
     1801 aggcattaga gttcttgaaa gatcgtccat tgtatgccga gaagtttgct ccttttgtta
     1861 aagaattagc ggtaatggtt gtgagatcac tggaaggcga agtattctcc tacccaaccg
     1921 tagaaactgt gcacaaggac aatatctgtc atattgtgta tgctccggcc agagttaatg
     1981 acaccatcca aaagaaagct caaatattag ctgaaaacac tgtgaagact ttcccaggcg
     2041 ctggaatctt cggagttgag atgttcctat tgtctgatgg agaacttctt gtaaatgaga
     2101 ttgctccaag gccccacaat tctggtcact atacaatcga tgcatgtgta acatctcagt
     2161 tcgaagcaca tgtaagagcc ataactggtc tgccaatgcc actagatttc accaaactat
     2221 ctacttccaa caccaacgct attatgctca atgttttggg tgctgaaaaa tctcacgggg
     2281 aattagagtt ttgtagaaga gccttagaaa cacccggtgc ttctgtatat ctgtacggaa
     2341 agaccacccg attggctcgt aagatgggtc atatcaacat aataggatct tccatgttgg
     2401 aagcagaaca aaagttagag tacattctag aagaatcaac ccacttacca tccagtactg
     2461 tatcagctga cactaaaccg ttggttggag ttatcatggg ttcagactct gatctacctg
     2521 tgatttcgaa aggttgcgat attttaaaac agtttggtgt tccattcgaa gttactattg
     2581 tctctgctca tagaacacca cagagaatga ccagatatgc ctttgaagcc gctagtagag
     2641 gtatcaaggc tatcattgca ggtgctggtg gtgctgctca tcttccagga atggttgctg
     2701 ccatgactcc gttgccagtc attggtgttc ctgtcaaggg ctctacgttg gatggtgtag
     2761 actcgctaca ctcgattgtc caaatgccta gaggtgttcc tgtggctacg gttgctatca
     2821 acaacgccac caatgccgct ctgttggcca tcaggatttt aggtacaatt gaccacaaat
     2881 ggcaaaagga aatgtccaag tatatgaatg caatggagac cgaagtgttg gggaaggcat
     2941 ccaacttgga atctgaaggg tatgaatcct atttgaagaa tcgtctttga atttagtatt
     3001 gttttttaat agatgtatat ataatagtac acgtaactta tctattccat tcataatttt
     3061 attttaaagg ttcggtagaa atttgtcctc caaaaagttg gttagagcct ggcagttttg
     3121 ataggcatta ttatagattg ggtaatattt accctgcacc tggaggaact ttgcaaagag
     3181 cctcatgtgc ggcgcgccag gccataatgg ccaaacggtt tctcaattac tatatactac
     3241 taaccattta cctgtagcgt atttcttttc cctcttcgcg aaagctcaag ggcatcttct
     3301 tgactcatga aaaatatctg gatttcttct gacagatcat cacccttgag cccaactctc
     3361 tagcctatga gtgtaagtga tagtcatctt gcaacagatt attttggaac gcaactaaca
     3421 aagcagatac acccttcagc agaatccttt ctggatattg tgaagaatga tcgccaaagt
     3481 cacagtcctg agacagttcc taatctttac cccatttaca agttcatcca atcagacttc
     3541 ttaacgcctc atctggctta tatcaagctt accaacagtt cagaaactcc cagtccaagt
     3601 ttcttgcttg aaagtgcgaa gaatggtgac accgttgaca ggtacacctt tatgggacat
     3661 tcccccagaa aaataatcaa gactgggcct ttagagggtg ctgaagttga ccccttggtg
     3721 cttctggaaa aagaactgaa gggcaccaga caagcgcaac ttcctggtat tcctcgtcta
     3781 agtggtggtg ccataggata catctcgtac gattgtatta agtactttga accaaaaact
     3841 gaaagaaaac tgaaagatgt tttgcaactt ccggaagcag ctttgatgtt gttcgacacg
     3901 atcgtggctt ttgacaatgt ttatcaaaga ttccaggtaa ttggaaacgt ttctctatcc
     3961 gttgatgact cggacgaagc tattcttgag aaatattata agacaagaga agaagtggaa
     4021 aagatcagta aagtggtatt tgacaataaa actgttccct actatgaaca gaaagatatt
     4081 attcaaggcc aaacgttcac ctctaatatt ggtcaggaag ggtatgaaaa ccatgttcgc
     4141 aagctgaaag aacatattct gaaaggagac atcttccaag ctgttccctc tcaaagggta
     4201 gccaggccga cctcattgca ccctttcaac atctatcgtc atttgagaac tgtcaatcct
     4261 tctccataca tgttctatat tgactatcta gacttccaag ttgttggtgc ttcacctgaa
     4321 ttactagtta aatccgacaa caacaacaaa atcatcacac atcctattgc tggaactctt
     4381 cccagaggta aaactatcga agaggacgac aattatgcta agcaattgaa gtcgtctttg
     4441 aaagacaggg ccgagcacgt catgctggta gatttggcca gaaatgatat taaccgtgtg
     4501 tgtgagccca ccagtaccac ggttgatcgt ttattgactg tggagagatt ttctcatgtg
     4561 atgcatcttg tgtcagaagt cagtggaaca ttgagaccaa acaagactcg cttcgatgct
     4621 ttcagatcca ttttcccagc aggaaccgtc tccggtgctc cgaaggtaag agcaatgcaa
     4681 ctcataggag aattggaagg agaaaagaga ggtgtttatg cgggggccgt aggacactgg
     4741 tcgtacgatg gaaaatcgat ggacacatgt attgccttaa gaacaatggt cgtcaaggac
     4801 ggtgtcgctt accttcaagc cggaggtgga attgtctacg attctgaccc ctatgacgag
     4861 tacatcgaaa ccatgaacaa aatgagatcc aacaataaca ccatcttgga ggctgagaaa
     4921 atctggaccg ataggttggc cagagacgag aatcaaagtg aatccgaaga aaacgatcaa
     4981 tgaacggagg acgtaagtag gaatttatgg tttggccata atggcctagc ttggcgtaat
     5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5881 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7201 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7261 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     7321 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7381 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7441 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7501 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7561 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     7621 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattg
//
