LOCUS       sequence-45430-e        6294 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 6294)
  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 6294)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2011-01-11"; volume: "108(2)"; pages: 
            "510-5. Epub 2010 Dec 22"
COMMENT     Sequence Label: pBaKdoO
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(878..1828)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1876..2098
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     3243..3260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3377..3398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3413..3418,3419..3436,3437..3443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3451..3467
                     /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     3456..3478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3475..3491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3475..3491
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3510..3530
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3512..3530
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4477..4493)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4478..4494)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4518..4537)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4519..4537)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4547..4564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4547..4563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4556..4578)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4705..5160
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4792..4811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5002..5023
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5186..5290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5291..5359,5360..6151)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5509..5528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tctctgttcg ggggttcact gacgctgcgt gactacgatg gtcaggttgc ggaggctatg
       61 gccctggtac gagcgctgaa caaaatgacg aaagcaggta tgcctgaaag cgtgcgtatt
      121 gcctgaaaac acaacccgct acgggggaga cttacccgaa atctgattta ttcaacaaag
      181 ccactttcag tatggatttg ggtgttacgg gggggggttg gtgcttaaaa tcgcgctgag
      241 gatttctgat ggtgttaagc gggcggtttt gagatgtaaa ctcgcccgtt taacataatg
      301 gatcttgcgc gcaccgcccg aacaccactc gccacaaaaa accgccggaa cgtccaaaag
      361 tacgggtttt gctgcccgca aacgggctgt tctggtgttg ctagtttgtt atcagaatcg
      421 cagatccggc ttcagccggt ttgccggctg aaagcgctat ttcttccaga attgccatga
      481 ttttttcccc acgggaggcg tcactggctc ccgtgttgtc ggcagctttg attcgataag
      541 cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa gttgtctcag
      601 gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc tattaggtgt
      661 tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc
      721 aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat
      781 ggacagtttt ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct
      841 tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg tatttagcca
      901 gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat
      961 catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg
     1021 cagttaaagc atcgtgtagt gtttttctta gtccgttatg taggtaggaa tctgatgtaa
     1081 tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga
     1141 gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct
     1201 tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa
     1261 cccattggtt aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa
     1321 attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta
     1381 ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat
     1441 tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta
     1501 attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt
     1561 taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta
     1621 atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga
     1681 acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac
     1741 agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat
     1801 ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac
     1861 tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg
     1921 gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg
     1981 taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata
     2041 gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac
     2101 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac
     2161 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc
     2221 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt
     2281 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt
     2341 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca
     2401 gggcgtttta tggcgggtct gcggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     2461 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     2521 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     2581 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     2641 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     2701 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     2761 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     2821 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     2881 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     2941 tcgccgaaaa tgacccagag cgctgccggc accctgtcct acgagttgca tgataaagaa
     3001 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3061 gttgaaggct ctcaagggca tcggcattcg ccattcaggc tgcctcctac caagacgact
     3121 tcaaatccga cccgtagaga tctcacatgt tctttcctgc gttatcccct gattctgtgg
     3181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3241 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3301 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3361 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     3421 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3481 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     3541 tggagctcca ccgcggtggc ggccgctcta gaaaggagat ataatgagcg aatcccagat
     3601 catcgaagtc ccgtccgccg actggagcgg acacaacctg tcggcgccgc gcgagcagct
     3661 gctggccgcg gtcgaggaag gcaaggtgct gtatttcccg cacctgcgct tcgcgatcga
     3721 aggcggcgag gaagcgctgc tcgatccgtc gctcgccgat ccgaaacgca agaacatcag
     3781 cctcgcgccg aacggcggcg tgctcgccgg cgtgctcggc gacagcgtca cgcagtcggc
     3841 cgtgcgcgcg ctcgtcgcgc gcttccagca gcaggccggc acgctcgtcg acggtctctt
     3901 tcccgaatac cgcggcaagc tgcgcgtcgc gccgacgagc ctgcggctga tgcaggtcga
     3961 gacgcgccag acgtcctggc gcaaggacga cagccggctg cacgtcgacg cgttcccgtc
     4021 gcggccgaac tacggcgagc ggatcctgcg tgtgttcacg aacgtgaacc cggccggcca
     4081 gccgcgcgtg tggcgcgtcg gcgagccgtt cgaggacgtc gcgaagcgct tcctgccgaa
     4141 gatccggccg cagttgccgg gctcggcgtg gctgctgaac ctgctgcacg tgacgaagtc
     4201 gccgcgcagc gcgtacgacc acctgatgct gaacctgcac gacgggatga aggctgatct
     4261 cgactaccag aagacctgtc cgcagcagac gatgccgttt ccgcccggca gcgtgtggat
     4321 atgtttctcg gatcagactt cgcacgctgt gatgtccggc cagttcatgc tcgagcagac
     4381 cttcttcctg ccggtcgacg cgatggtgcg gcgcgaatgc gcgccgctgg gaattctcga
     4441 gcgcctgacc ggccgggcgc tggtttgaaa gcttatcgat accgtcgacc tcgagggggg
     4501 gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     4561 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4621 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4681 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4741 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4801 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4861 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4921 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4981 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5041 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5101 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5161 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5221 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5281 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5341 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5401 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5461 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5521 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5581 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5641 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5701 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5761 aactcgcctt gatcgctggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5821 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5881 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5941 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6001 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6061 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6121 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6181 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6241 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt caga
//
