LOCUS       40924_30655        8050 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 8050)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8050)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8050
                     /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             1539..1580
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1590..1607
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     promoter        2715..3090
                     /label=ADE2 promoter
     CDS             3091..4803
                     /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(6022..6610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6784..7641)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7642..7746)
                     /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 tcgagaagag agaggctgaa gcatcgatga attcacgtgg cccagccggc cgtggatcca
     1501 cgcgtcgtcg acccgcggcg gccgccagct tactagtagg taagcctatc cctaaccctc
     1561 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga tctagtatac
     1621 aattctaggg ctgcctgttt ggatattttt ataatttttg agagtttgcc aactaatgtt
     1681 tttctcttct atgatattta tcatgtagtt ggatatcttt ttttctttga aaagttttta
     1741 tttagttctt tctctctttt ttataatgtt ttcagtttct ttcctttata ttgaaaatac
     1801 gatgttcttt gtctttcttt tttaaatcct caattataat gtgtaccgtt tgatcaactt
     1861 gtaaaattgt ttgtagtcct gtaatcttaa ttgacgattg gttgatagtt taccccagta
     1921 tattgtgcgg ctgagatatt acccgattcg gaagattgat tttttcaaca gacataaacc
     1981 cataggagat ttagtacagc gacatatgct aattgcctga taatgctcac taacattttc
     2041 tccttttaat ctggtatatg catacaggta tgttgatatg gccttgttga tcaaagctac
     2101 gaaagtgtag actatttccc aaacagcccc caaccaccac ctcacacgcg cacactgcga
     2161 agtgaaagcg ctcccttatt ttcttctccg ctggttcaaa acaaaaaaga atctaatctg
     2221 tagacacacg ccccgtaaaa cccaatcttc atcatcgtaa ttttcagtga cggacgatcg
     2281 atcaaccata tagtttgtta gtgactgagc ttcagtagtt gacctcttga agaaataacg
     2341 tcttgaacaa catactttat acctaccaac attcgccatc aacaacagtg tgacactgac
     2401 ataactttac atccattcat cgacaaaaaa atgtctctag ttcggattca tgcttatggg
     2461 ttagctattt cgcccaatgt gtccatctga cattactatt ttgcatttta atttaattag
     2521 aacttgacta gcgcactacc agcatatcat ctcatttccg taaataccaa atgtattata
     2581 tattgaaagc ttttgaccag gttattataa aagaaacttc atgctcgaaa aagatcattt
     2641 cgaaaagttg cctagtttca tgaaatttta aagcagttta tataaatttt accttttgat
     2701 gcggaattga ctttttcttg aataatacat aacttttctt aaaagaatca aagacagata
     2761 aaatttaaga gatattaaat attagtgaga agccgagaat tttgtaacac caacataaca
     2821 ctgacatctt taacaacttt taattatgat acatttctta cgtcatgatt gattattaca
     2881 gctatgctga caaatgactc ttgttgcatg gctacgaacc gggtaatact aagtgattga
     2941 ctcttgctga ccttttatta agaactaaat ggacaatatt atggagcatt tcatgtataa
     3001 attggtgcgt aaaatcgttg gatctctctt ctaagtacat cctactataa caatcaagaa
     3061 aaacaagaaa atcggacaaa acaatcaagt atggattcta gaacagttgg tatattagga
     3121 gggggacaat tgggacgtat gattgttgag gcagcaaaca ggctcaacat taagacggta
     3181 atactagatg ctgaaaattc tcctgccaaa caaataagca actccaatga ccacgttaat
     3241 ggctcctttt ccaatcctct tgatatcgaa aaactagctg aaaaatgtga tgtgctaacg
     3301 attgagattg agcatgttga tgttcctaca ctaaagaatc ttcaagtaaa acatcccaaa
     3361 ttaaaaattt acccttctcc agaaacaatc agattgatac aagacaaata tattcaaaaa
     3421 gagcatttaa tcaaaaatgg tatagcagtt acccaaagtg ttcctgtgga acaagccagt
     3481 gagacgtccc tattgaatgt tggaagagat ttgggttttc cattcgtctt gaagtcgagg
     3541 actttggcat acgatggaag aggtaacttc gttgtaaaga ataaggaaat gattccggaa
     3601 gctttggaag tactgaagga tcgtcctttg tacgccgaaa aatgggcacc atttactaaa
     3661 gaattagcag tcatgattgt gagatctgtt aacggtttag tgttttctta cccaattgta
     3721 gagactatcc acaaggacaa tatttgtgac ttatgttatg cgcctgctag agttccggac
     3781 tccgttcaac ttaaggcgaa gttgttggca gaaaatgcaa tcaaatcttt tcccggttgt
     3841 ggtatatttg gtgtggaaat gttctattta gaaacagggg aattgcttat taacgaaatt
     3901 gccccaaggc ctcacaactc tggacattat accattgatg cttgcgtcac ttctcaattt
     3961 gaagctcatt tgagatcaat attggatttg ccaatgccaa agaatttcac atctttctcc
     4021 accattacaa cgaacgccat tatgctaaat gttcttggag acaaacatac aaaagataaa
     4081 gagctagaaa cttgcgaaag agcattggcg actccaggtt cctcagtgta cttatatgga
     4141 aaagagtcta gacctaacag aaaagtaggt cacataaata ttattgcctc cagtatggcg
     4201 gaatgtgaac aaaggctgaa ctacattaca ggtagaactg atattccaat caaaatctct
     4261 gtcgctcaaa agttggactt ggaagcaatg gtcaaaccat tggttggaat catcatggga
     4321 tcagactctg acttgccggt aatgtctgcc gcatgtgcgg ttttaaaaga ttttggcgtt
     4381 ccatttgaag tgacaatagt ctctgctcat agaactccac ataggatgtc agcatatgct
     4441 atttccgcaa gcaagcgtgg aattaaaaca attatcgctg gagctggtgg ggctgctcac
     4501 ttgccaggta tggtggctgc aatgacacca cttcctgtca tcggtgtgcc cgtaaaaggt
     4561 tcttgtctag atggagtaga ttctttacat tcaattgtgc aaatgcctag aggtgttcca
     4621 gtagctaccg tcgctattaa taatagtacg aacgctgcgc tgttggctgt cagactgctt
     4681 ggcgcttatg attcaagtta tacaacgaaa atggaacagt ttttattaaa gcaagaagaa
     4741 gaagttcttg tcaaagcaca aaagttagaa actgtcggtt acgaagctta tctagaaaac
     4801 aagtaatata taagtttatt gatatacttg tacagcaaat aattataaaa tgatatacct
     4861 attttttagg ctttgttatg attacatcaa atgtggactt catacataga aatcaacgct
     4921 tacaggtgtc cttttttaag aatttcatac ataagatcta ctagaatcaa tgttaaacct
     4981 gttatcatta gaacagccat ctccagaagg ttctatccaa gtgaaaactt tggtcttcaa
     5041 gccaaagact gctaaaacac aaaccccagt cccaattatt gttgttgctg aaacttctac
     5101 ttctactccg ggtggtgtta tagccaaagc ttcgggcact aaagagccaa gattagcaaa
     5161 cgatgaattg accaaccaat tctttggttc tcctgctaaa gagttttcta ttgccagctt
     5221 atcaaacgaa cacaaagacg gtaaagtcaa gatcattatt gatgctggga tcaaagatga
     5281 tgactcgact aattacgtta tttccaacac tgttgctaag ttagttttaa ccggtaagca
     5341 attattaggt ttcttatcca actacgaacc aaccatcgtt gattttactg ctgaagctgc
     5401 aaccgctgcc ccaactccag ttgttaagaa agaacctgct gcatctggta agcttgatga
     5461 tgccaaatta attggtgtta ctgctgacaa ggaaaaagat ttctccaact ggtactccca
     5521 aattctaatg aagggtgaaa tgttagacta ctacgatgtc tctggttgtt acattatgag
     5581 accagcttct tacacaattt gggaaaagat tcaatcttgg ttcgattcga agattaaaga
     5641 agatggtgtt actaatgctt atttcccaat gtttgtctca tccaaggttt tagaaaagga
     5701 aaaggaccac atcgatggtt tcgctcctga agttgcttgg gttaccagag ctggtaaatc
     5761 tgaattagaa gaaccaattg ctattagacc aacttctgaa actgttatgg gtaccctgca
     5821 ggggccggcc ttaattaagg cgcgcccttc ctcgctcact gactcgctgc gctcggtcgt
     5881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7741 tccgcgcaca tttccccgaa aagtgccacc tgtatgcggt gtgaaatacc gcacagatgc
     7801 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     7861 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     7921 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     7981 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat caagggcgat
     8041 ggcccactac
//
