LOCUS       sequence-448140-        5439 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 5439)
  AUTHORS   Chung HS, Raetz CR
  TITLE     Interchangeable domains in the Kdo transferases of Escherichia coli 
            and Haemophilus influenzae.
  JOURNAL   Biochemistry. 2010 May 18;49(19):4126-37.
  PUBMED    20394418
REFERENCE   2  (bases 1 to 5439)
  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: 
            "Biochemistry."; date: "2010-05-18"; volume: "49(19)"; pages: 
            "4126-37"
COMMENT     Sequence Label: pBAD33.1
FEATURES             Location/Qualifiers
     source          1..5439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        310..412
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             413..1072
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(479..498)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(1628..2083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1765..1786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1977..1996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(2541..2632)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2651..2678)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2770..2856)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3009..3026
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3092..3124)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             3132..3154
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3214..3498)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(3272..3291)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3525..4403
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(4583..4602)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      4678..5223
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5124..5143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5340..5357
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg
       61 gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg acagctgata
      121 gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca gtaagttggc
      181 agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc acttcgcaga
      241 ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact tttggcgaaa
      301 atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
      361 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
      421 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      481 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      541 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      601 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
      661 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      721 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
      781 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      841 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      901 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
      961 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     1021 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     1081 aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg
     1141 gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg gcagggtcgt
     1201 taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg aagcagtgtg
     1261 accgtgtgct tctcaaatgc ctgactggta acaggattag cagagcgagg tatgtaggcg
     1321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     1381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1621 tccttttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1681 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1741 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1801 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1861 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1921 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1981 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2041 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaatcaa tctaaagtat
     2101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2401 ttcgccagtt aatagtttgc tgcacccaac tgatcttcag catcttttac tttcaccagc
     2461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aaagagtttg
     2641 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt
     2701 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc
     2761 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     2821 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     2881 catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     2941 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc
     3001 tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccaa
     3061 gcttgtcgac ggagctcgaa ttcggatccc gacccatttg ctgtccacca gtcatgctag
     3121 ccatatgtat atctccttct taaagttaaa caaaattatt tctagaggat ccccgggtac
     3181 cgagctcgaa ttcgctagcc caaaaaaacg ggtatggaga aacagtagag agttgcgata
     3241 aaaagcgtca ggtaggatcc gctaatctta tggataaaaa tgctatggca tagcaaagtg
     3301 tgacgccgtg caaataatca atgtggactt ttctgccgtg attatagaca cttttgttac
     3361 gcgtttttgt catggctttg gtcccgcttt gttacagaat gcttttaata agcggggtta
     3421 ccggtttggt tagcgagaag agccagtaaa agacgcagtg acggcaatgt ctgatgcaat
     3481 atggacaatt ggtttcttct ctgaatggcg ggagtatgaa aagtatggct gaagcgcaaa
     3541 atgatcccct gctgccggga tactcgttta atgcccatct ggtggcgggt ttaacgccga
     3601 ttgaggccaa cggttatctc gattttttta tcgaccgacc gctgggaatg aaaggttata
     3661 ttctcaatct caccattcgc ggtcaggggg tggtgaaaaa tcagggacga gaatttgttt
     3721 gccgaccggg tgatattttg ctgttcccgc caggagagat tcatcactac ggtcgtcatc
     3781 cggaggctcg cgaatggtat caccagtggg tttactttcg tccgcgcgcc tactggcatg
     3841 aatggcttaa ctggccgtca atatttgcca atacggggtt ctttcgcccg gatgaagcgc
     3901 accagccgca tttcagcgac ctgtttgggc aaatcattaa cgccgggcaa ggggaagggc
     3961 gctattcgga gctgctggcg ataaatctgc ttgagcaatt gttactgcgg cgcatggaag
     4021 cgattaacga gtcgctccat ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca
     4081 tcagcgatca cctggcagac agcaattttg atatcgccag cgtcgcacag catgtttgct
     4141 tgtcgccgtc gcgtctgtca catcttttcc gccagcagtt agggattagc gtcttaagct
     4201 ggcgcgagga ccaacgtatc agccaggcga agctgctttt gagcaccacc cggatgccta
     4261 tcgccaccgt cggtcgcaat gttggttttg acgatcaact ctatttctcg cgggtattta
     4321 aaaaatgcac cggggccagc ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg
     4381 atgtagccgt caagttgtca taattggtaa cgaatcagac aattgacggc ttgacggagt
     4441 agcatagggt ttgcagaatc cctgcttcgt ccatttgaca ggcacattat gcatcgatga
     4501 taagctgtca aacatgagca gatcctctac gccggacgca tcgtggccgg catcaccggc
     4561 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
     4621 cgccacttcg ggctcatgag cctagaaata ttttatctga ttaataagat gatcttcttg
     4681 agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg
     4741 cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg
     4801 agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac
     4861 taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg
     4921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt
     4981 cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa
     5041 tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga
     5101 gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5161 cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg
     5221 aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca
     5281 ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta
     5341 gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg
     5401 gtgcagcctt ttttctcctg ccacatgaag cacttcact
//
