LOCUS       40924_44834        8510 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 8510)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8510)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..1227
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             1302..4346
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPVPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     CDS             4374..4415
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             4425..4442
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    4471..4695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4741..5169
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5183..5513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        5561..5608
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             5627..6022
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6183..6316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6353..6369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6377..6393)
                     /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(6401..6431)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6446..6467)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6755..7343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7517..8374)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8375..8479)
                     /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 atcgaaacga tgatagatcc cgtcgtttta caacgtcgtg
     1321 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1381 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1441 atggcgaatg gcgctttgcc tggtttccgg taccagaagc ggtgccggaa agctggctgg
     1501 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
     1561 acgatgcgcc catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc
     1621 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
     1681 aggaaggcca gacgcgaatt atttttgatg gcgttaactc ggcgtttcat ctgtggtgca
     1741 acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc gtctgaattt gacctgagcg
     1801 catttttacg cgccggagaa aaccgcctcg cggtgatggt gctgcgttgg agtgacggca
     1861 gttatctgga agatcaggat atgtggcgga tgagcggcat tttccgtgac gtctcgttgc
     1921 tgcataaacc gactacacaa atcagcgatt tccatgttgc cactcgcttt aatgatgatt
     1981 tcagccgcgc tgtactggag gctgaagttc agatgtgcgg cgagttgcgt gactacctac
     2041 gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg
     2101 gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg
     2161 tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac
     2221 tgcacaccgc cgacggcacg ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg
     2281 tgcggattga aaatggtctg ctgctgctga acggcaagcc gttgctgatt cgaggcgtta
     2341 accgtcacga gcatcatcct ctgcatggtc aggtcatgga tgagcagacg atggtgcagg
     2401 atatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat tatccgaacc
     2461 atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat gaagccaata
     2521 ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg
     2581 cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg agtgtgatca
     2641 tctggtcgct ggggaatgaa tcaggccacg gcgctaatca cgacgcgctg tatcgctgga
     2701 tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg cggcggagcc gacaccacgg
     2761 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctg
     2821 tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg agagacgcgc ccgctgatcc
     2881 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
     2941 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     3001 tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat tttggcgata
     3061 cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc
     3121 cagcgctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta tccgggcaaa
     3181 ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc ctgcactgga
     3241 tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat gtcgctccac
     3301 aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc gggcaactct
     3361 ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc gggcacatca
     3421 gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc
     3481 acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc
     3541 gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac
     3601 aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg
     3661 taagtgaagc gacccgcatt gaccctaacg cctgggtcga acgctggaag gcggcgggcc
     3721 attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct gatgcggtgc
     3781 tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc agccggaaaa
     3841 cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa gtggcgagcg
     3901 atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta gcagagcggg
     3961 taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact gccgcctgtt
     4021 ttgaccgctg ggatctgcca ttgtcagaca tgtatacccc gtacgtcttc ccgagcgaaa
     4081 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
     4141 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
     4201 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
     4261 gcgacgactc ctggagcccg tcagtatcgg cggaattcca gctgagcgcc ggtcgctacc
     4321 attaccagtt ggtctggtgt caaaaagcgg ccgctcgagg tcacccattc gaaggtaagc
     4381 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     4441 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4681 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     4741 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4801 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4861 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     4921 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4981 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5041 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     5101 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5161 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     5221 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5281 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5341 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5401 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     5461 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5521 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     5581 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
     5641 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5701 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5761 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5821 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5881 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5941 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     6001 ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc
     6061 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6121 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6181 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6241 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6301 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     6961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7621 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8461 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
