LOCUS       40924_44849        5459 bp DNA     circular SYN 18-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5459)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5459)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..1227
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             1323..1364
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1374..1391
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    1420..1644
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1690..2118
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2132..2462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2510..2557
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2576..2971
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3132..3265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3302..3318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3326..3342)
                     /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(3350..3380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3395..3416)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3704..4292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4466..5323)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5324..5428)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
       61 cctcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
      121 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
      181 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
      241 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
      301 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
      361 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
      421 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
      481 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
      541 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
      601 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
      661 aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat gggctggggc
      721 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
      781 ttgcgggggc ggcagttatg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
      841 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
      901 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
      961 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     1021 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     1081 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     1141 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     1201 tggccgtttt tggctttttt gttagacgaa gcttggtacc gagctcggat ccactagtcc
     1261 agtgtggtgg aattctgcag atatccagca cagtggcggc cgctcgaggt cacccattcg
     1321 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     1381 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     1441 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1501 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1561 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1621 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1681 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1741 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1801 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     1861 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1921 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1981 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2041 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     2101 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2161 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2221 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2281 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2341 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2401 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2461 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     2521 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     2581 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     2641 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     2701 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     2761 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     2821 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     2881 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     2941 attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag gagcaggact
     3001 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3061 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3121 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3241 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3301 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3361 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3421 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3541 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3601 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3661 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3721 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3781 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3841 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     3901 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3961 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4021 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4081 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4141 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4201 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4261 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4321 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4381 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4441 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5341 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5401 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
