LOCUS       sequence-7448-ex        6058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  AUTHORS   Ku MJ, Yoon HJ, Ahn HJ, Kim HW, Baek SH, Suh SW
  TITLE     Crystallization and preliminary X-ray crystallographic studies of 
            3-deoxy-manno-octulosonate cytidylyltransferase from Haemophilus 
            influenzae.
  JOURNAL   Acta Crystallogr D Biol Crystallogr. 2003 Jan . 59(Pt 1):180-2.
  PUBMED    12499564
REFERENCE   2  (bases 1 to 6058)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Acta 
            Crystallogr D Biol Crystallogr. 2003 Jan . 59(Pt 1):180-2."
COMMENT     Sequence Label: HI0058/pET-21a(+)c-tag
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(932..949)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(959..976)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             995..1017
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1032..1056
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     complement(1056..1075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1057..1075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1159..1178)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1384..1461
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1462..2544
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1481..1500)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3353..3544
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3538..3560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3646..3788
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3701..3720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3803..3820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3974..4562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4054..4073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4673..4692)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4684..5499
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4684..5499
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4757..4776)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5592..6047)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5729..5750)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5941..5960
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagatgtcat ttaccgtgat
      181 tatccccgct cgttttgcat caagtcgttt gccaggaaaa cctcttgctg atattaaagg
      241 taagccaatg attcaacatg tatttgagaa agcactgcag tctggggcga gccgagtgat
      301 tattgccacc gataatgaaa atgtggctga tgttgccaaa agttttggtg ctgaagtctg
      361 tatgacttcg gttaatcata attctggtac agaacgttta gcagaagttg tagaaaaatt
      421 agctattcct gacaatgaaa tcattgtcaa tattcaaggc gatgagcctt tgattcctcc
      481 cgttatcgtg cgacaagtgg cagataattt agcaaaattt aatgtcaata tggcaagcct
      541 tgcggtaaaa attcacgatg ctgaggaatt atttaatcca aatgcagtga aagtattaac
      601 agataaagat ggctatgtgc tgtatttttc ccgttcggtt attccttatg atcgtgatca
      661 gtttatgaat ttgcaggatg ttcagaaagt acagctttct gacgcttact tacgtcatat
      721 tggcatttac gcatatcgtg cgggtttcat aaaacaatat gtgcaatggg caccgactca
      781 acttgaaaat ctagaaaaac ttgagcagct tcgtgtgtta tataacggcg aacgtattca
      841 cgtagaactt gcgaaagaag tgcctgcagt gggggtggat accgccgaag atttggaaaa
      901 agtgcgggca attttagcgg cgaattaaat ggctgccgcg cggcaccagg ccgctgctgt
      961 gatgatgatg atgatggctg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt
     1021 atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc
     1081 gcgggatcga gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     1141 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     1201 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     1261 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     1321 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
     1381 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1441 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1501 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     1561 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     1621 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     1681 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1741 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1801 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1861 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1921 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1981 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     2041 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     2101 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     2161 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     2221 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     2281 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2341 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2401 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2461 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2521 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     2581 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2641 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     2701 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     2761 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     2821 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     2881 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2941 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     3001 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     3061 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     3121 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     3181 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     3241 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     3301 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     3361 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     3421 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     3481 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3541 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3601 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     3661 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     3721 ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3781 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4381 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4621 aagggatttt ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     4681 gttatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg
     4741 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     4801 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     4861 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     4921 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     4981 gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     5041 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     5101 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     5161 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     5221 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     5281 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     5341 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     5401 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     5461 aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata
     5521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5581 agtgccacct gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5641 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5701 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5761 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5821 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5881 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5941 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     6001 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
