LOCUS       40924_15125        6305 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 6305)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6305)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      751..938
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(950..966)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1402..2748)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2862..3662)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3663..3878)
                     /label=URA3 promoter
     promoter        3989..4093
                     /label=AmpR promoter
     CDS             4094..4951
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5125..5713
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6001..6022
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6037..6067
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6075..6091
                     /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)."
     primer_bind     6099..6115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggagaaactc ctgcgagaat ttcgtaaact cgcgcgcatt gcatctgtat ttcctaatgc
       61 ggcacttcca ggcctcgatc gagaccgttt atccattgct tttttgttgt ctttttccct
      121 cgttcacaga aagtctgaag aagctatagt agaactatga gctttttttg tttctgtttt
      181 cctttttttt ttttttacct ctgtggaaat tgttactctc acactcttta gttcgtttgt
      241 ttgttttgtt tattccaatt atgaccggtg acgaaacgtg gtcgatggtg ggtaccgctt
      301 atgctcccct ccattagttt cgattatata aaaaggccaa atattgtatt attttcaaat
      361 gtcctatcat tatcgtctaa catctaattt ctcttaaatt ttttctcttt ctttcctata
      421 acaccaatag tgaaaatctt tttttcttct atatctacaa aaactttttt tttctatcaa
      481 cctcgttgat aaattttttc tttaacaatc gttaataatt aattaattgg aaaataacca
      541 ttttttctct cttttataca cacattcaaa agaaagaaaa aaaatatacc ccagcctcga
      601 gcggccgcgg atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag
      661 gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt
      721 ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt
      781 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
      841 aaacgaraat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
      901 tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg
      961 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1021 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1081 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     1141 gcggtatttc acaccgcata atcggatcgt acttgttacc catcattgaa ttttgaacat
     1201 ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata
     1261 gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct
     1321 attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca
     1381 cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     1441 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     1501 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     1561 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     1621 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     1681 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     1741 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     1801 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     1861 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     1921 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     1981 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     2041 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     2101 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     2161 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     2221 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     2281 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     2341 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     2401 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
     2461 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
     2521 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
     2581 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
     2641 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
     2701 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga tccgatgata
     2761 agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt
     2821 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     2881 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     2941 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     3001 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     3061 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     3121 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     3181 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     3241 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     3301 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     3361 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     3421 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     3481 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     3541 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     3601 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     3661 atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa gaatactggg
     3721 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     3781 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga
     3841 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa
     3901 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4801 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5041 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5101 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5161 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5221 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5281 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5341 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5401 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5461 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5521 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5581 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5641 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5701 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5761 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5821 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5881 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5941 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6001 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6061 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6121 ttacgccaag cttcctgaaa cggagaaaca taaacaggca ttgctgggat cacccataca
     6181 tcactctgtt ttgcctgacc ttttccggta atttgaaaac aaacccggtc tcgaagcgga
     6241 gatccggcga taattaccgc agaaataaac ccatacacga gacgtagaac cagccgcaca
     6301 tggcc
//
