LOCUS       40924_2812        6412 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 6412)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6412)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..416
                     /label=URA3 promoter
     CDS             417..1217
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2014..2261)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2264..2280
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2314..2330)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2345..2786)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(2810..2828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2849..2865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2873..2889)
                     /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(2897..2927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2942..2963)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4013..4870)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4871..4975)
                     /label=AmpR promoter
     rep_origin      complement(5002..6344)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg gccgcaaatt aaagccttcg agcgtcccaa
     2041 aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa
     2101 aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata
     2161 aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg
     2221 gggagggcgt gaatgtaagc gtgacataac taattacatg actcgaggtc gacggtatcg
     2281 ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga atccggggtt
     2341 ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga tacttttatt
     2401 acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta aagtggttat
     2461 gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg ctgattaatt
     2521 accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt taatttgatt
     2581 cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc ataaccataa
     2641 aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca catctgcgtt
     2701 tcaggaacgc gaccggtgaa gacgaggacg cacggaggag agtcttcctt cggagggctg
     2761 tcacccgctc ggcggcttct aatccgtact agagctccag cttttgttcc ctttagtgag
     2821 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2881 cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct
     2941 aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3001 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3061 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4981 ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc
     5041 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     5101 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     5161 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     5221 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     5281 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     5341 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     5401 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     5461 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     5521 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     5581 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     5641 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     5701 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     5761 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     5821 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     5881 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     5941 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     6001 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     6061 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     6121 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     6181 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     6241 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     6301 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt
     6361 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
