LOCUS       40924_5749        7115 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 7115)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7115)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             332..349
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             353..385
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             389..412
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             431..3475
                     /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"
     terminator      3685..3771
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3863..3890
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3910..4001
                     /label=AmpR promoter
     CDS             4002..4859
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5033..5621
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5807..5947)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6214..7089)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctaacaggag gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag
      361 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccaat
      421 gatagatccc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
      481 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
      541 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggt
      601 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
      661 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtaaccta
      721 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
      781 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
      841 cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag
      901 tcgtttgccg tctgaatttg acctgagcgc atttttacgc gccggagaaa accgcctcgc
      961 ggtgatggtg ctgcgttgga gtgacggcag ttatctggaa gatcaggata tgtggcggat
     1021 gagcggcatt ttccgtgacg tctcgttgct gcataaaccg actacacaaa tcagcgattt
     1081 ccatgttgcc actcgcttta atgatgattt cagccgcgct gtactggagg ctgaagttca
     1141 gatgtgcggc gagttgcgtg actacctacg ggtaacagtt tctttatggc agggtgaaac
     1201 gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta
     1261 tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat
     1321 cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc tgattgaagc
     1381 agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa
     1441 cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag catcatcctc tgcatggtca
     1501 ggtcatggat gagcagacga tggtgcagga tatcctgctg atgaagcaga acaactttaa
     1561 cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg tacacgctgt gcgaccgcta
     1621 cggcctgtat gtggtggatg aagccaatat tgaaacccac ggcatggtgc caatgaatcg
     1681 tctgaccgat gatccgcgct ggctaccggc gatgagcgaa cgcgtaacgc gaatggtgca
     1741 gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg gggaatgaat caggccacgg
     1801 cgctaatcac gacgcgctgt atcgctggat caaatctgtc gatccttccc gcccggtgca
     1861 gtatgaaggc ggcggagccg acaccacggc caccgatatt atttgcccga tgtacgcgcg
     1921 cgtggatgaa gaccagccct tcccggctgt gccgaaatgg tccatcaaaa aatggctttc
     1981 gctacctgga gagacgcgcc cgctgatcct ttgcgaatac gcccacgcga tgggtaacag
     2041 tcttggcggt ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg
     2101 cttcgtctgg gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg
     2161 gtcggcttac ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg
     2221 tctggtcttt gccgaccgca cgccgcatcc agcgctgacg gaagcaaaac accagcagca
     2281 gtttttccag ttccgtttat ccgggcaaac catcgaagtg accagcgaat acctgttccg
     2341 tcatagcgat aacgagctcc tgcactggat ggtggcgctg gatggtaagc cgctggcaag
     2401 cggtgaagtg cctctggatg tcgctccaca aggtaaacag ttgattgaac tgcctgaact
     2461 accgcagccg gagagcgccg ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc
     2521 gaccgcatgg tcagaagccg ggcacatcag cgcctggcag cagtggcgtc tggcggaaaa
     2581 cctcagtgtg acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat
     2641 ggatttttgc atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct
     2701 ttcacagatg tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac
     2761 ccgtgcaccg ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc
     2821 ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg
     2881 cacggcagat acacttgctg atgcggtgct gattacgacc gctcacgcgt ggcagcatca
     2941 ggggaaaacc ttatttatca gccggaaaac ctaccggatt gatggtagtg gtcaaatggc
     3001 gattaccgtt gatgttgaag tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa
     3061 ctgccagctg gcgcaggtag cagagcgggt aaactggctc ggattagggc cgcaagaaaa
     3121 ctatcccgac cgccttactg ccgcctgttt tgaccgctgg gatctgccat tgtcagacat
     3181 gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc tgcgggacgc gcgaattgaa
     3241 ttatggccca caccagtggc gcggcgactt ccagttcaac atcagccgct acagtcaaca
     3301 gcaactgatg gaaaccagcc atcgccatct gctgcacgcg gaagaaggca catggctgaa
     3361 tatcgacggt ttccatatgg ggattggtgg cgacgactcc tggagcccgt cagtatcggc
     3421 ggaattccag ctgagcgccg gtcgctacca ttaccagttg gtctggtgtc aaaaataagc
     3481 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     3541 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     3601 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     3661 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3721 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     3781 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     3841 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     3901 caaactcttt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3961 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4021 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4081 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4141 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4201 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     4261 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4321 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4381 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4441 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4501 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4561 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4621 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4681 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4741 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4801 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4861 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4921 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4981 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5041 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5101 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5161 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5221 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5281 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5341 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5401 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5461 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5521 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5581 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5641 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5701 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5761 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     5821 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     5881 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     5941 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6001 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6061 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca
     6121 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     6181 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     6241 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     6301 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     6361 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     6421 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
     6481 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     6541 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     6601 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     6661 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     6721 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     6781 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     6841 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     6901 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     6961 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     7021 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     7081 ttcagccata cttttcatac tcccgccatt cagag
//
