LOCUS       40924_40087        8138 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 8138)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8138)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        258..478
                     /label=URA3 promoter
     rep_origin      complement(1417..1872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2017..2033
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2043..2061
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2774..3181)
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(3210..3226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(3236..3483)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3804..4832)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(4930..4946)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4961..5402)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(5426..5444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5465..5481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5489..5505)
                     /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(5513..5543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5558..5579)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5867..6455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6629..7486)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7487..7591)
                     /label=AmpR promoter
     misc_feature    7628..8131
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
       61 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
      121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      241 gcaccatacc acagcttttc aattcaattc atcatttttt ttttattctt ttttttgatt
      301 tcggtttctt tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
      361 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
      421 cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat
      481 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
      541 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
      601 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
      661 tgtggatatc ttgactgatt tttccatgca tggagggcac agttaagccg ctaaaggcat
      721 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
      781 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
      841 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
      901 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
      961 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     1021 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     1081 tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg
     1141 tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     1201 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     1261 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     1321 actcacaaat tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata
     1381 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     1441 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1501 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1561 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1621 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1681 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1741 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1801 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1861 tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1921 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1981 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     2041 cgtaatacga ctcactatag ggcgaattgg gcgtacacgc gtctgtacag aaaaaaaaga
     2101 aaaatttgaa atataaataa cgttcttaat actaacataa ctataaaaaa ataaataggg
     2161 acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt ggggggaggg
     2221 cgtgaatgta agcgtgacat aactaattac atgatatcga caaaggaaaa gggggacgga
     2281 tccgtcgagt gggtggtgag cagctcgccg tcgcgcagca ggcgccagcg ctccatccac
     2341 gcagtgacct caggcggaaa acgggaagac acactcatga gatgcctgca agcaattcgt
     2401 tctgtatcag gcgcaggagc gtcccgtccg ggtcgaccaa agcggccatc gtgcctcccc
     2461 actcctgcag ttcgggggca tggatgcgcg gatagccgct gctggtttcc tggatgccga
     2521 cggatttgca ctgccggtag aactccgcga ggtcgtccag cctcaggcag cagctgaacc
     2581 aactcgcgag gggatcgagc ccggggtggg cgaagaactc cagcatgaga tccccgcgct
     2641 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     2701 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     2761 attcgagctc gcccttagat tagattgcta tgctttcttt ctaatgagca agaagtaaaa
     2821 aaagttgtaa tagaacaaga aaaatgaaac tgaaacttga gaaattgaag accgtttatt
     2881 aacttaaata tcaatgggag gtcatcgaaa gagaaaaaaa tcaaaaaaaa aaattttcaa
     2941 gaaaaagaaa cgtgataaaa atttttattg cctttttcga cgaagaaaaa gaaacgaggc
     3001 ggtctctttt ttcttttcca aacctttagt acgggtaatt aacgacaccc tagaggaaga
     3061 aagaggggaa atttagtatg ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga
     3121 gtccgagaaa atctggaaga gtaaaaaagg agtagaaaca ttttgaagct atggtgtgtg
     3181 ggggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg cggccgcaaa
     3241 ttaaagcctt cgagcgtccc aaaaccttct caagcaaggt tttcagtata atgttacatg
     3301 cgtacacgcg tctgtacaga aaaaaaagaa aaatttgaaa tataaataac gttcttaata
     3361 ctaacataac tataaaaaaa taaataggga cctagacttc aggttgtcta actccttcct
     3421 tttcggttag agcggatgtg gggggagggc gtgaatgtaa gcgtgacata actaattaca
     3481 tgactcgagg tcgacaattc caaccttacc caagagttcg ccaaactcag acatcacttt
     3541 agcaaaaccg cgccgtgctt cttcctcggt ggcattcatc acgaaatgtt cagcactacg
     3601 catacttttg acaggaaacg caacggatat tgagtcaata tcaggcattc tatcgctcag
     3661 ctttacagtg acaatgacgg ctggcgactg aatattagtg cttacagaca gcactacata
     3721 ttttccgtcg atgttgaaat cctttctcat atgtcaccat aaatatcaaa taattatagc
     3781 aatcatttac gcgttaatgg ctaatcgcca tcttccagca ggcgcaccat tgcccctgtt
     3841 tcactatcca gggtacggat atagttcatg acaatattta cattggtcca gccaccagct
     3901 tgcatgatct ccggtattga aactccagcg cgggccatat ctcgcgcggc tccgacacgg
     3961 gcactgtgtc cagaccaggc caggtatctc tgaccagagt catccttagc gccgtaaatc
     4021 aatcgatgag ttgcttcaaa aatcccttcc agggcgcgag ttgatagctg gctggtggca
     4081 gatggcgcgg caacaccatt ttttctgacc cggcaaaaca ggtagttatt cggatcatca
     4141 gctacaccag agacggaaat ccatcgctcg accagtttag ttacccccag gctaagtgcc
     4201 ttctctacac ctgcggtgct aaccagcgtt ttcgttctgc caatatggat taacattctc
     4261 ccaccgtcag tacgtgagat atctttaacc ctgatcctgg caatttcggc tatacgtaac
     4321 agggtgttat aagcaatccc cagaaatgcc agattacgta tatcctggca gcgatcgcta
     4381 ttttccatga gtgaacgaac ctggtcgaaa tcagtgcgtt cgaacgctag agcctgtttt
     4441 gcacgttcac cggcatcaac gttttctttt cggatccgcc gcataaccag tgaaacagca
     4501 ttgctgtcac ttggtcgtgg cagcccggac cgacgatgaa gcatgtttag ctggcccaaa
     4561 tgttgctgga tagtttttac tgccagaccg cgcgcctgaa gatatagaag ataatcgcga
     4621 acatcttcag gttctgcggg aaaccatttc cggttattca acttgcacca tgccgcccac
     4681 gaccggcaaa cggacagaag cattttccag gtatgctcag aaaacgcctg gcgatccctg
     4741 aacatgtcca tcaggttctt gcgaacctca tcactcgttg catcgaccgg taatgcaggc
     4801 aaattttggt gtacggtcag taaattggac atttaacact cagataatgg ttttaagtaa
     4861 agtgtacagg atcggctctg cccctcgacg gtatcgataa gcttgatatc gaattcctgc
     4921 agcccggggg atccactagt tctagaatcc ggggtttttt ctccttgacg ttaaagtata
     4981 gaggtatatt aacaattttt tgttgatact tttattacat ttgaataaga agtaatacaa
     5041 accgaaaatg ttgaaagtat tagttaaagt ggttatgcag tttttgcatt tatatatctg
     5101 ttaatagatc aaaaatcatc gcttcgctga ttaattaccc cagaaataag gctaaaaaac
     5161 taatcgcatt atcatcctat ggttgttaat ttgattcgtt catttgaagg tttgtggggc
     5221 caggttactg ccaatttttc ctcttcataa ccataaaagc tagtattgta gaatctttat
     5281 tgttcggagc agtgcggcgc gaggcacatc tgcgtttcag gaacgcgacc ggtgaagacg
     5341 aggacgcacg gaggagagtc ttccttcgga gggctgtcac ccgctcggcg gcttctaatc
     5401 cgtactagag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     5461 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag
     5521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa
     5581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6301 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     7621 ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa
     7681 attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat
     7741 gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct
     7801 caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct
     7861 gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg
     7921 tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt
     7981 tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac
     8041 aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata
     8101 cgaggcgcgt gtaagttaca ggcaagcgat ccgtccta
//
