LOCUS       40924_40352        4885 bp DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4885)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4885)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(470..710)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        complement(841..859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(871..889)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(909..925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(933..949)
                     /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(957..987)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1002..1023)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1311..1899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2073..2930)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     protein_bind    2992..3008
                     /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     3016..3032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(3069..3202)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3335..4357)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(4414..4624)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttttccaa aaaactaccg
      421 ttgttatagg tgtctcttga acacctataa caacggtagt ggatcccgcg tcctttccac
      481 aagatatata aacccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc
      541 attttaaaac ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac
      601 gtattttgta ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca
      661 gccttgtatc gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg
      721 accttggcgc gcgctcggcg cgcggtcacg ctccgtcacg tggtgcgttt tgcctgcgcg
      781 tctttccact ggggaattca tgcttctcct ccctttagtg agggtaattc tctctctctc
      841 cctatagtga gtcgtattaa ttccttctct tctatagtgt cacctaaatc gttgcaattc
      901 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      961 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1021 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1081 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1141 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1201 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1261 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1321 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1381 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1441 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1501 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1561 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1621 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1681 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1741 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1801 agagttggta gctcttgatc cggcaaaaaa accaccgctg gtagcggtgg tttttttgtt
     1861 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1921 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1981 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2041 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2101 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2161 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2221 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2281 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2341 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2401 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2461 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2521 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2581 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2641 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2701 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2761 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2821 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2881 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2941 tttcaatatt caggcttacc tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     3001 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg
     3061 tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3121 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3181 tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct
     3241 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     3301 cggcggtgga atcgaaatct cgtagcacgt gctattcctt tgccctcgga cgagtgctgg
     3361 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc
     3421 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
     3481 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
     3541 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat
     3601 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
     3661 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
     3721 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
     3781 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
     3841 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     3901 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     3961 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct
     4021 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     4081 gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg
     4141 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     4201 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     4261 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     4321 tctcgacaga cgtcgcggtg agttcaggct ttttcatcac gtgctgatca gatccgaaaa
     4381 tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcccc
     4441 actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa
     4501 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     4561 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     4621 ctgctgggga gcctggggac tttccacacc tggttgctaa ctaattgaga tgcatgcttt
     4681 gcatacttct gcctgctgga atattattga agcatttatc agggttattg tctcatgagc
     4741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4801 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4861 aggcgtatca cgaggccctt tcgtc
//
