LOCUS       40924_44844        5467 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 5467)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5467)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..1227
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             1331..1372
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1382..1399
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    1428..1652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1698..2126
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2140..2470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2518..2565
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2584..2979
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3140..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3334..3350)
                     /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(3358..3388)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3403..3424)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3712..4300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4474..5331)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5332..5436)
                     /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 cgctcgagtc tagagggccc
     1321 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     1381 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     1441 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1501 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1561 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1621 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1681 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1741 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1801 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     1861 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1921 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1981 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2041 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     2101 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2161 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     2221 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2281 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2341 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2401 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2461 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     2521 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     2581 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     2641 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     2701 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     2761 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     2821 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     2881 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     2941 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga
     3001 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3061 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3121 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3241 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3301 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3361 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3421 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3481 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3901 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4141 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5461 tgacgtc
//
