LOCUS       sequence-45429-e        6312 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 6312)
  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 6312)
  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: pYpKdoO
FEATURES             Location/Qualifiers
     source          1..6312
                     /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(4495..4511)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4496..4512)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4536..4555)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4537..4555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4565..4582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4565..4581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4574..4596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4723..5178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4810..4829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5020..5041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5204..5308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5309..5377,5378..6169)
                     /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(5527..5546)
                     /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 atattgatgt gtattgataa
     3601 accagagtca gatccgctga tccttacact tcctaacacg cgctgggacc agaatgcaga
     3661 ggataatacc ggtgccattg aggcacttga gcaaggtaaa gtcctgttct tacctcacct
     3721 gacttttgaa ctgagtgagc aggaaaaaca gttactggac cctaccctgg ttgatgttaa
     3781 acgcaagaat atcagtttca aaccgttaga aggtgtatta actggcgtga ctgatgcaag
     3841 caaaatcgca ttaacccgcc agttattgga tcgttattat caaagttgcc tcagtttggt
     3901 acagcaactc ctcccggtct atacgcatgc gctacacagc ccgaccaata gcttgagatt
     3961 acaccctgtg gccgcctggc gcgacagtac gtcatggcgt aaagatgata gccgcttaca
     4021 tgtagatgct ttcccgtcac gcccaaacta cggtgagcgt atcatccgca tattcaccaa
     4081 tattaacccg catggtgaag cgcgttcctg gcgtgtcggt gaagatttta cacaacttgc
     4141 cagccgctat ttgccacaac tggatagcta ttcatcgctc agcagttggt tgcaacataa
     4201 aatcggtatc accaagcgcc cccgtagcca ttacgaccat ctgatgctcc aactgcatga
     4261 caaaatgaag gccgatctcg cttatcaaaa agatggctta caacaggcca ttgagttccc
     4321 accaggcagc agttggatat gcttttcgga tcaaacacct catgcggcga tgggcggaca
     4381 atttatgttg gagcaaacgt tactactacc tgttgaagga atgaaatcct cccagcgctc
     4441 accactgaaa atactggaag cgttgacaca aaaaacgctg atataaaagc ttatcgatac
     4501 cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     4561 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     4621 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     4681 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4741 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4801 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4861 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4921 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4981 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5041 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5101 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5161 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5221 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5281 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5341 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5401 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5461 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5521 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5581 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5641 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5701 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5761 caacatgggg gatcatgtaa ctcgccttga tcgctgggaa ccggagctga atgaagccat
     5821 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5881 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5941 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6001 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6061 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6121 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6181 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6241 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6301 ctgagcgtca ga
//
