LOCUS       40924_37958        5388 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 5388)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5388)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..348
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             355..1374
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     rep_origin      complement(1663..2118)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2263..2279
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2289..2307
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2316..2423)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2436..2454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2475..2491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2499..2515)
                     /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(2523..2553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2568..2589)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2877..3465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3639..4496)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4497..4601)
                     /label=AmpR promoter
     misc_feature    4638..5141
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
       61 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
      121 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcggacctg
      181 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
      241 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
      301 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaaaag
      361 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
      421 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
      481 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
      541 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
      601 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
      661 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
      721 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
      781 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
      841 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
      901 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
      961 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     1021 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     1081 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     1141 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     1201 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     1261 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     1321 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataatct
     1381 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     1441 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     1501 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     1561 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acggcggtgt
     1621 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1681 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1741 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1801 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1861 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1921 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1981 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2041 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2101 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     2161 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     2221 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2281 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     2341 acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact agttctagag
     2401 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2521 cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa
     2581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3301 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4621 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
     4681 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     4741 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     4801 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     4861 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     4921 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     4981 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     5041 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     5101 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt
     5161 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5221 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5281 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5341 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtg
//
