LOCUS       sequence-45431-e        6287 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  AUTHORS   Chung HS, Raetz CR
  TITLE     Dioxygenases in Burkholderia ambifaria and Yersinia pestis that 
            hydroxylate the outer Kdo unit of lipopolysaccharide.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Jan 11;108(2):510-5. Epub 2010 Dec 
            22.
  PUBMED    21178073
REFERENCE   2  (bases 1 to 6287)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2011-01-11"; volume: "108(2)"; pages: 
            "510-5. Epub 2010 Dec 22"
COMMENT     Sequence Label: pYpKdoO.1
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(98..117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     308..329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2403..2545
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2471..2490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2631..2653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2647..2838)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3647..4729)
                     /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     4691..4710
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4730..4807)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5013..5032
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5116..5135
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5116..5134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5135..5159
                     /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)."
     RBS             5174..5196
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6131..6148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6201..6219)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6215..6262
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
       61 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      121 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      181 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      421 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt
     2461 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     2521 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     2581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     2701 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     2761 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2821 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2881 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2941 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3001 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     3061 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     3121 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     3181 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     3241 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     3301 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     3361 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     3421 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     3481 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     3541 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     3601 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3721 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     3781 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     3841 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     3901 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     3961 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     4021 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     4081 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     4141 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     4201 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     4261 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     4321 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     4381 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     4441 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     4501 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     4561 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     4621 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     4681 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     4741 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     4801 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     4861 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     4921 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     4981 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     5041 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     5101 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     5161 ctctagaaat aattttgttt aactttaaga aggagatata catatgttga tgtgtattga
     5221 taaaccagag tcagatccgc tgatccttac acttcctaac acgcgctggg accagaatgc
     5281 agaggataat accggtgcca ttgaggcact tgagcaaggt aaagtcctgt tcttacctca
     5341 cctgactttt gaactgagtg agcaggaaaa acagttactg gaccctaccc tggttgatgt
     5401 taaacgcaag aatatcagtt tcaaaccgtt agaaggtgta ttaactggcg tgactgatgc
     5461 aagcaaaatc gcattaaccc gccagttatt ggatcgttat tatcaaagtt gcctcagttt
     5521 ggtacagcaa ctcctcccgg tctatacgca tgcgctacac agcccgacca atagcttgag
     5581 attacaccct gtggccgcct ggcgcgacag tacgtcatgg cgtaaagatg atagccgctt
     5641 acatgtagat gctttcccgt cacgcccaaa ctacggtgag cgtatcatcc gcatattcac
     5701 caatattaac ccgcatggtg aagcgcgttc ctggcgtgtc ggtgaagatt ttacacaact
     5761 tgccagccgc tatttgccac aactggatag ctattcatcg ctcagcagtt ggttgcaaca
     5821 taaaatcggt atcaccaagc gcccccgtag ccattacgac catctgatgc tccaactgca
     5881 tgacaaaatg aaggccgatc tcgcttatca aaaagatggc ttacaacagg ccattgagtt
     5941 cccaccaggc agcagttgga tatgcttttc ggatcaaaca cctcatgcgg cgatgggcgg
     6001 acaatttatg ttggagcaaa cgttactact acctgttgaa ggaatgaaat cctcccagcg
     6061 ctcaccactg aaaatactgg aagcgttgac acaaaaaacg ctgatataaa agcttgcggc
     6121 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     6181 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     6241 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggat
//
