LOCUS       40924_30645        8045 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8045)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8045)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1197..1463
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1534..1575
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1585..1602
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     promoter        2710..3085
                     /label=ADE2 promoter
     CDS             3086..4798
                     /codon_start=1
                     /label=ADE2
                     /note="phosphoribosylaminoimidazole carboxylase, required
                     for adenine biosynthesis"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(6017..6605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6779..7636)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7637..7741)
                     /label=AmpR promoter
ORIGIN
        1 ggcgcgcctt aattaaggcc ggcccctgca gggggccccg agatggtaca tacttaaaag
       61 ctgccatatt gaggaacttc aaagttttat ctgtttttag aattaaaaga cgattgttgt
      121 aacaaaacgt tgtgcctaca taaactcaaa ttaatggaaa tagcctgttt tgaaaaatac
      181 accttcttaa gtactgacaa agttttgtta aatgactatc gaacaagcca tgaaatagca
      241 catttctgcc agtcactttt aacactttcc tgcttgctgg ttgactctcc tcatacaaac
      301 acccaaaagg gaaactttca gtgtggggac acttgacatc tcacatgcac cccagattaa
      361 tttccccaga cgatgcggag acaagacaaa acaacccttt gtcctgctct tttctttctc
      421 acaccgcgtg ggtgtgtgcg caggcaggca ggcaggcagc gggctgcctg ccatttctaa
      481 tcgctgctcc tcccccctgg cttcaaataa cagcctgctg ctatctgtga ccagattggg
      541 acacccccct cccctccgaa tgatccatca ccttttgtcg tactccgaca atgatccttc
      601 cctgtcatct tctggcaatc agctccttca ataattaaat caaataagca taaatagtaa
      661 aatcgcatac aaacgtcatg aaaagtttta tctctatggc caacggatag tctatctgct
      721 taattccatc cactttggga accgttctct ctttacccca gattctcaaa gctaatatct
      781 gccccttgtc tattgtcctt tctccgtgta caagcggagc ttttgcctcc catcctcttg
      841 ctttgtttcg gttatttttt ttttcttttg aaactcttgg tcaaatcaaa tcaaacaaaa
      901 ccaaaccttc tattccatca gatcaacctt gttcaacatt ctataaatcg atataaatat
      961 aaccttatcc ctcccttgtt ttttaccaat taatcaatct tcaaatttca aatattttct
     1021 acttgcttta ttactcagta ttaacatttg tttaaaccaa ctataacttt taactggctt
     1081 tagaagtttt atttaacatc agtttcaatt tacatcttta tttattaacg aaatctttac
     1141 gaattaactc aatcaaaact tttacgaaaa aaaaatctta ctattaagag ctcaaaatga
     1201 gatttccttc aatttttact gctgttttat tcgcagcatc ctccgcatta gctgctccag
     1261 tcaacactac aacagaagat gaaacggcac aaattccggc tgaagctgtc atcggttact
     1321 cagatttaga aggggatttc gatgttgctg ttttgccatt ttccaacagc acaaataacg
     1381 ggttattgtt tataaatact actattgcca gcattgctgc taaagaagaa ggggtatctc
     1441 tcgagaaaag agaggctgaa gctgaattca cgtggcccag ccggccgtgg atccacgcgt
     1501 cgtcgacccg cggcggccgc cagcttacta gtaggtaagc ctatccctaa ccctctcctc
     1561 ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgatctag tatacaattc
     1621 tagggctgcc tgtttggata tttttataat ttttgagagt ttgccaacta atgtttttct
     1681 cttctatgat atttatcatg tagttggata tctttttttc tttgaaaagt ttttatttag
     1741 ttctttctct cttttttata atgttttcag tttctttcct ttatattgaa aatacgatgt
     1801 tctttgtctt tcttttttaa atcctcaatt ataatgtgta ccgtttgatc aacttgtaaa
     1861 attgtttgta gtcctgtaat cttaattgac gattggttga tagtttaccc cagtatattg
     1921 tgcggctgag atattacccg attcggaaga ttgatttttt caacagacat aaacccatag
     1981 gagatttagt acagcgacat atgctaattg cctgataatg ctcactaaca ttttctcctt
     2041 ttaatctggt atatgcatac aggtatgttg atatggcctt gttgatcaaa gctacgaaag
     2101 tgtagactat ttcccaaaca gcccccaacc accacctcac acgcgcacac tgcgaagtga
     2161 aagcgctccc ttattttctt ctccgctggt tcaaaacaaa aaagaatcta atctgtagac
     2221 acacgccccg taaaacccaa tcttcatcat cgtaattttc agtgacggac gatcgatcaa
     2281 ccatatagtt tgttagtgac tgagcttcag tagttgacct cttgaagaaa taacgtcttg
     2341 aacaacatac tttataccta ccaacattcg ccatcaacaa cagtgtgaca ctgacataac
     2401 tttacatcca ttcatcgaca aaaaaatgtc tctagttcgg attcatgctt atgggttagc
     2461 tatttcgccc aatgtgtcca tctgacatta ctattttgca ttttaattta attagaactt
     2521 gactagcgca ctaccagcat atcatctcat ttccgtaaat accaaatgta ttatatattg
     2581 aaagcttttg accaggttat tataaaagaa acttcatgct cgaaaaagat catttcgaaa
     2641 agttgcctag tttcatgaaa ttttaaagca gtttatataa attttacctt ttgatgcgga
     2701 attgactttt tcttgaataa tacataactt ttcttaaaag aatcaaagac agataaaatt
     2761 taagagatat taaatattag tgagaagccg agaattttgt aacaccaaca taacactgac
     2821 atctttaaca acttttaatt atgatacatt tcttacgtca tgattgatta ttacagctat
     2881 gctgacaaat gactcttgtt gcatggctac gaaccgggta atactaagtg attgactctt
     2941 gctgaccttt tattaagaac taaatggaca atattatgga gcatttcatg tataaattgg
     3001 tgcgtaaaat cgttggatct ctcttctaag tacatcctac tataacaatc aagaaaaaca
     3061 agaaaatcgg acaaaacaat caagtatgga ttctagaaca gttggtatat taggaggggg
     3121 acaattggga cgtatgattg ttgaggcagc aaacaggctc aacattaaga cggtaatact
     3181 agatgctgaa aattctcctg ccaaacaaat aagcaactcc aatgaccacg ttaatggctc
     3241 cttttccaat cctcttgata tcgaaaaact agctgaaaaa tgtgatgtgc taacgattga
     3301 gattgagcat gttgatgttc ctacactaaa gaatcttcaa gtaaaacatc ccaaattaaa
     3361 aatttaccct tctccagaaa caatcagatt gatacaagac aaatatattc aaaaagagca
     3421 tttaatcaaa aatggtatag cagttaccca aagtgttcct gtggaacaag ccagtgagac
     3481 gtccctattg aatgttggaa gagatttggg ttttccattc gtcttgaagt cgaggacttt
     3541 ggcatacgat ggaagaggta acttcgttgt aaagaataag gaaatgattc cggaagcttt
     3601 ggaagtactg aaggatcgtc ctttgtacgc cgaaaaatgg gcaccattta ctaaagaatt
     3661 agcagtcatg attgtgagat ctgttaacgg tttagtgttt tcttacccaa ttgtagagac
     3721 tatccacaag gacaatattt gtgacttatg ttatgcgcct gctagagttc cggactccgt
     3781 tcaacttaag gcgaagttgt tggcagaaaa tgcaatcaaa tcttttcccg gttgtggtat
     3841 atttggtgtg gaaatgttct atttagaaac aggggaattg cttattaacg aaattgcccc
     3901 aaggcctcac aactctggac attataccat tgatgcttgc gtcacttctc aatttgaagc
     3961 tcatttgaga tcaatattgg atttgccaat gccaaagaat ttcacatctt tctccaccat
     4021 tacaacgaac gccattatgc taaatgttct tggagacaaa catacaaaag ataaagagct
     4081 agaaacttgc gaaagagcat tggcgactcc aggttcctca gtgtacttat atggaaaaga
     4141 gtctagacct aacagaaaag taggtcacat aaatattatt gcctccagta tggcggaatg
     4201 tgaacaaagg ctgaactaca ttacaggtag aactgatatt ccaatcaaaa tctctgtcgc
     4261 tcaaaagttg gacttggaag caatggtcaa accattggtt ggaatcatca tgggatcaga
     4321 ctctgacttg ccggtaatgt ctgccgcatg tgcggtttta aaagattttg gcgttccatt
     4381 tgaagtgaca atagtctctg ctcatagaac tccacatagg atgtcagcat atgctatttc
     4441 cgcaagcaag cgtggaatta aaacaattat cgctggagct ggtggggctg ctcacttgcc
     4501 aggtatggtg gctgcaatga caccacttcc tgtcatcggt gtgcccgtaa aaggttcttg
     4561 tctagatgga gtagattctt tacattcaat tgtgcaaatg cctagaggtg ttccagtagc
     4621 taccgtcgct attaataata gtacgaacgc tgcgctgttg gctgtcagac tgcttggcgc
     4681 ttatgattca agttatacaa cgaaaatgga acagttttta ttaaagcaag aagaagaagt
     4741 tcttgtcaaa gcacaaaagt tagaaactgt cggttacgaa gcttatctag aaaacaagta
     4801 atatataagt ttattgatat acttgtacag caaataatta taaaatgata tacctatttt
     4861 ttaggctttg ttatgattac atcaaatgtg gacttcatac atagaaatca acgcttacag
     4921 gtgtcctttt ttaagaattt catacataag atctactaga atcaatgtta aacctgttat
     4981 cattagaaca gccatctcca gaaggttcta tccaagtgaa aactttggtc ttcaagccaa
     5041 agactgctaa aacacaaacc ccagtcccaa ttattgttgt tgctgaaact tctacttcta
     5101 ctccgggtgg tgttatagcc aaagcttcgg gcactaaaga gccaagatta gcaaacgatg
     5161 aattgaccaa ccaattcttt ggttctcctg ctaaagagtt ttctattgcc agcttatcaa
     5221 acgaacacaa agacggtaaa gtcaagatca ttattgatgc tgggatcaaa gatgatgact
     5281 cgactaatta cgttatttcc aacactgttg ctaagttagt tttaaccggt aagcaattat
     5341 taggtttctt atccaactac gaaccaacca tcgttgattt tactgctgaa gctgcaaccg
     5401 ctgccccaac tccagttgtt aagaaagaac ctgctgcatc tggtaagctt gatgatgcca
     5461 aattaattgg tgttactgct gacaaggaaa aagatttctc caactggtac tcccaaattc
     5521 taatgaaggg tgaaatgtta gactactacg atgtctctgg ttgttacatt atgagaccag
     5581 cttcttacac aatttgggaa aagattcaat cttggttcga ttcgaagatt aaagaagatg
     5641 gtgttactaa tgcttatttc ccaatgtttg tctcatccaa ggttttagaa aaggaaaagg
     5701 accacatcga tggtttcgct cctgaagttg cttgggttac cagagctggt aaatctgaat
     5761 tagaagaacc aattgctatt agaccaactt ctgaaactgt tatgggtacc ctgcaggggc
     5821 cggccttaat taaggcgcgc ccttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7741 gcacatttcc ccgaaaagtg ccacctgtat gcggtgtgaa ataccgcaca gatgcgtaag
     7801 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7861 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7921 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     7981 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaagg gcgatggccc
     8041 actac
//
