LOCUS       40924_30650        8049 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 8049)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8049)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8049
                     /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             1538..1579
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1589..1606
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     promoter        2714..3089
                     /label=ADE2 promoter
     CDS             3090..4802
                     /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(6021..6609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6783..7640)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7641..7745)
                     /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 gctgcaggaa ttcacgtggc ccagccggcc gtggatccac
     1501 gcgtcgtcga cccgcggcgg ccgccagctt actagtaggt aagcctatcc ctaaccctct
     1561 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgat ctagtataca
     1621 attctagggc tgcctgtttg gatattttta taatttttga gagtttgcca actaatgttt
     1681 ttctcttcta tgatatttat catgtagttg gatatctttt tttctttgaa aagtttttat
     1741 ttagttcttt ctctcttttt tataatgttt tcagtttctt tcctttatat tgaaaatacg
     1801 atgttctttg tctttctttt ttaaatcctc aattataatg tgtaccgttt gatcaacttg
     1861 taaaattgtt tgtagtcctg taatcttaat tgacgattgg ttgatagttt accccagtat
     1921 attgtgcggc tgagatatta cccgattcgg aagattgatt ttttcaacag acataaaccc
     1981 ataggagatt tagtacagcg acatatgcta attgcctgat aatgctcact aacattttct
     2041 ccttttaatc tggtatatgc atacaggtat gttgatatgg ccttgttgat caaagctacg
     2101 aaagtgtaga ctatttccca aacagccccc aaccaccacc tcacacgcgc acactgcgaa
     2161 gtgaaagcgc tcccttattt tcttctccgc tggttcaaaa caaaaaagaa tctaatctgt
     2221 agacacacgc cccgtaaaac ccaatcttca tcatcgtaat tttcagtgac ggacgatcga
     2281 tcaaccatat agtttgttag tgactgagct tcagtagttg acctcttgaa gaaataacgt
     2341 cttgaacaac atactttata cctaccaaca ttcgccatca acaacagtgt gacactgaca
     2401 taactttaca tccattcatc gacaaaaaaa tgtctctagt tcggattcat gcttatgggt
     2461 tagctatttc gcccaatgtg tccatctgac attactattt tgcattttaa tttaattaga
     2521 acttgactag cgcactacca gcatatcatc tcatttccgt aaataccaaa tgtattatat
     2581 attgaaagct tttgaccagg ttattataaa agaaacttca tgctcgaaaa agatcatttc
     2641 gaaaagttgc ctagtttcat gaaattttaa agcagtttat ataaatttta ccttttgatg
     2701 cggaattgac tttttcttga ataatacata acttttctta aaagaatcaa agacagataa
     2761 aatttaagag atattaaata ttagtgagaa gccgagaatt ttgtaacacc aacataacac
     2821 tgacatcttt aacaactttt aattatgata catttcttac gtcatgattg attattacag
     2881 ctatgctgac aaatgactct tgttgcatgg ctacgaaccg ggtaatacta agtgattgac
     2941 tcttgctgac cttttattaa gaactaaatg gacaatatta tggagcattt catgtataaa
     3001 ttggtgcgta aaatcgttgg atctctcttc taagtacatc ctactataac aatcaagaaa
     3061 aacaagaaaa tcggacaaaa caatcaagta tggattctag aacagttggt atattaggag
     3121 ggggacaatt gggacgtatg attgttgagg cagcaaacag gctcaacatt aagacggtaa
     3181 tactagatgc tgaaaattct cctgccaaac aaataagcaa ctccaatgac cacgttaatg
     3241 gctccttttc caatcctctt gatatcgaaa aactagctga aaaatgtgat gtgctaacga
     3301 ttgagattga gcatgttgat gttcctacac taaagaatct tcaagtaaaa catcccaaat
     3361 taaaaattta cccttctcca gaaacaatca gattgataca agacaaatat attcaaaaag
     3421 agcatttaat caaaaatggt atagcagtta cccaaagtgt tcctgtggaa caagccagtg
     3481 agacgtccct attgaatgtt ggaagagatt tgggttttcc attcgtcttg aagtcgagga
     3541 ctttggcata cgatggaaga ggtaacttcg ttgtaaagaa taaggaaatg attccggaag
     3601 ctttggaagt actgaaggat cgtcctttgt acgccgaaaa atgggcacca tttactaaag
     3661 aattagcagt catgattgtg agatctgtta acggtttagt gttttcttac ccaattgtag
     3721 agactatcca caaggacaat atttgtgact tatgttatgc gcctgctaga gttccggact
     3781 ccgttcaact taaggcgaag ttgttggcag aaaatgcaat caaatctttt cccggttgtg
     3841 gtatatttgg tgtggaaatg ttctatttag aaacagggga attgcttatt aacgaaattg
     3901 ccccaaggcc tcacaactct ggacattata ccattgatgc ttgcgtcact tctcaatttg
     3961 aagctcattt gagatcaata ttggatttgc caatgccaaa gaatttcaca tctttctcca
     4021 ccattacaac gaacgccatt atgctaaatg ttcttggaga caaacataca aaagataaag
     4081 agctagaaac ttgcgaaaga gcattggcga ctccaggttc ctcagtgtac ttatatggaa
     4141 aagagtctag acctaacaga aaagtaggtc acataaatat tattgcctcc agtatggcgg
     4201 aatgtgaaca aaggctgaac tacattacag gtagaactga tattccaatc aaaatctctg
     4261 tcgctcaaaa gttggacttg gaagcaatgg tcaaaccatt ggttggaatc atcatgggat
     4321 cagactctga cttgccggta atgtctgccg catgtgcggt tttaaaagat tttggcgttc
     4381 catttgaagt gacaatagtc tctgctcata gaactccaca taggatgtca gcatatgcta
     4441 tttccgcaag caagcgtgga attaaaacaa ttatcgctgg agctggtggg gctgctcact
     4501 tgccaggtat ggtggctgca atgacaccac ttcctgtcat cggtgtgccc gtaaaaggtt
     4561 cttgtctaga tggagtagat tctttacatt caattgtgca aatgcctaga ggtgttccag
     4621 tagctaccgt cgctattaat aatagtacga acgctgcgct gttggctgtc agactgcttg
     4681 gcgcttatga ttcaagttat acaacgaaaa tggaacagtt tttattaaag caagaagaag
     4741 aagttcttgt caaagcacaa aagttagaaa ctgtcggtta cgaagcttat ctagaaaaca
     4801 agtaatatat aagtttattg atatacttgt acagcaaata attataaaat gatataccta
     4861 ttttttaggc tttgttatga ttacatcaaa tgtggacttc atacatagaa atcaacgctt
     4921 acaggtgtcc ttttttaaga atttcataca taagatctac tagaatcaat gttaaacctg
     4981 ttatcattag aacagccatc tccagaaggt tctatccaag tgaaaacttt ggtcttcaag
     5041 ccaaagactg ctaaaacaca aaccccagtc ccaattattg ttgttgctga aacttctact
     5101 tctactccgg gtggtgttat agccaaagct tcgggcacta aagagccaag attagcaaac
     5161 gatgaattga ccaaccaatt ctttggttct cctgctaaag agttttctat tgccagctta
     5221 tcaaacgaac acaaagacgg taaagtcaag atcattattg atgctgggat caaagatgat
     5281 gactcgacta attacgttat ttccaacact gttgctaagt tagttttaac cggtaagcaa
     5341 ttattaggtt tcttatccaa ctacgaacca accatcgttg attttactgc tgaagctgca
     5401 accgctgccc caactccagt tgttaagaaa gaacctgctg catctggtaa gcttgatgat
     5461 gccaaattaa ttggtgttac tgctgacaag gaaaaagatt tctccaactg gtactcccaa
     5521 attctaatga agggtgaaat gttagactac tacgatgtct ctggttgtta cattatgaga
     5581 ccagcttctt acacaatttg ggaaaagatt caatcttggt tcgattcgaa gattaaagaa
     5641 gatggtgtta ctaatgctta tttcccaatg tttgtctcat ccaaggtttt agaaaaggaa
     5701 aaggaccaca tcgatggttt cgctcctgaa gttgcttggg ttaccagagc tggtaaatct
     5761 gaattagaag aaccaattgc tattagacca acttctgaaa ctgttatggg taccctgcag
     5821 gggccggcct taattaaggc gcgcccttcc tcgctcactg actcgctgcg ctcggtcgtt
     5881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6421 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7081 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7381 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7741 ccgcgcacat ttccccgaaa agtgccacct gtatgcggtg tgaaataccg cacagatgcg
     7801 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     7861 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     7921 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     7981 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc aagggcgatg
     8041 gcccactac
//
