LOCUS       Exported                5438 bp DNA     circular SYN 15-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5438)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5438)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5438)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..819)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      941..1529
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1715..1854)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1959..2147)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3708..3726
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3758..3780
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3793..3810
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             3814..4914
                     /codon_start=1
                     /label=MBP
                     /note="maltose binding protein from E. coli"
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             4966..4986
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             5054..5071
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      5138..5185
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
       61 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      121 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      181 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      241 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
      301 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      361 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
      421 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
      481 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
      541 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
      601 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
      661 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
      721 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
      781 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
      841 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
      901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
      961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1081 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1561 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1621 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1681 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1741 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     1801 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     1861 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     1921 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1981 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     2041 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     2101 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     2161 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     2221 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     2281 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     2341 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     2401 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     2461 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     2521 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     2581 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     2641 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     2701 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     2761 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     2821 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     2881 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     2941 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     3001 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     3061 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     3121 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3181 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     3241 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     3301 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     3361 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct
     3421 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     3481 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     3541 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     3601 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     3661 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     3721 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     3781 tataccatga aacatcacca tcaccatcac cccatgaaaa tcgaagaagg taaactggta
     3841 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
     3901 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
     3961 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
     4021 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
     4081 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
     4141 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
     4201 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
     4261 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
     4321 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
     4381 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
     4441 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
     4501 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
     4561 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
     4621 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
     4681 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     4741 gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca gaaaggtgaa
     4801 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     4861 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg
     4921 agctcgaaca acaacaacaa taacaataac aacaacccca tgagcgagaa tctttatttt
     4981 cagggcgcca tggactaatg aggtaccgga tccgaattcg agctccgtcg acaagcttgc
     5041 ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     5101 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5161 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atccacagga
     5221 cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga
     5281 ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca
     5341 tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga
     5401 tatcccgcaa gaggcccggc agtaccggca taaccaag
//
