LOCUS       sequence-297712-       10324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli plasmid that contains the expression optimized genes for 
            BB0323, P13, DipA and Lmp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10324)
  AUTHORS   Robertson KE, Truong CD, Craciunescu FM, Chiu PL, Fromme P, Hansen 
            DT
  TITLE     Membrane directed expression in Escherichia coli of BBA57 and other 
            virulence factors from the Lyme disease agent Borrelia burgdorferi.
  JOURNAL   Sci Rep. 2019 Nov 26;9(1):17606. doi: 10.1038/s41598-019-53830-x.
  PUBMED    31772280
REFERENCE   2  (bases 1 to 10324)
  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: "Sci Rep."; 
            date: "2019-11-26"; volume: "9(1):17606. doi"; pages: " 
            10.1038/s41598-019-53830-x"
COMMENT     Sequence Label: pCC1-4k-BBclone2
FEATURES             Location/Qualifiers
     source          1..10324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6217..6239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6228..6244
                     /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)."
     promoter        complement(join(6252..6258,6259..6276,6277..6282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6297..6318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(6538..7197)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7112..7131
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7198..7300)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      8158..8772
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      8249..8780
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      8848..9067
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             9158..9913
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    9916..10166
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
ORIGIN
        1 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtgaattg gcgaattcat gaacatcaaa aacaagctga tcagcctgct
      121 gattgtggtt gcgatcagct tcattgcgtg caagaccccg ccggagagcc gtgaaagcaa
      181 aaacgcgaag atcgcgcagc cggacaacaa aaactttcaa ctgcgtgaca tcaaagatat
      241 taagaacgaa ctgattcgtg agcgtggcca cctgttctat agcaaagaat ttaacgaggc
      301 ggaacgtctg gaggaagcga tgaagcagag cttcagcaag aaaaaggcga tcgagggtaa
      361 cgaaattgcg ctgaaagtgc tggagcgtta caagaccatc attcgtgaga cccgtgagaa
      421 gaaggagaag accaactacc tgaaagaaaa catcgagaag tatctgaacg acgcggaagc
      481 gaacgaggcg tacatctgga ttccgctgga gattgatgag gttaacaacc tgtatttcga
      541 ggcgacccgt aaatacaaga actatgacct ggataacgcg ctggacatgt acagcaaagc
      601 gtttaaccgt gcgcagcaag cggcgaaaaa cgcgaaggaa gcgaaagcgc tgaaggaaac
      661 cgatgagcgt atgtacaaac agctgaaggc gctggaggcg gcgagcaacc tgccgatcta
      721 tagcaacaac aaactgatta agccgagccc gtggaacggc cgtgcgttca tcaaagaacg
      781 taacagccac ctgaacctgc tgaacaccaa caaggacacc tacctgctgg gcgaggcgga
      841 aattagcatc ccgattgtgc tggcgtatga ggaaaaagtt gaaatcgcga aaaacagcaa
      901 gccgcaagag caatttaaga ccctggaact gattgagcgt agccgtaccc tgtgggaaaa
      961 aggtgtggag gcgaaaaacg ttaagaactt ccgtctggcg aacgaactgt ttctggagag
     1021 cgcgcgttac ctggaagcgt atcagagcaa cgcgagcagc gagctgtacg ttatcaagat
     1081 tggcaacacc ctgtggggta tcagcaaaaa gctgtacaac gacccgtatc tgtggccgaa
     1141 gatttggttc gcgaaccgtc agaaaatcca aaacccggat ctgattcaca gcaactggaa
     1201 gatcattatc ccggcgaaaa tgaacaagct gctgatcttc gtgctggcga ccttttgcgt
     1261 tttcagcagc tttgcgcagg cgaacgacag caaaaacggc gcgtttggta tgagcgcggg
     1321 tgaaaagctg ctggtgtacg agaccagcaa acaggacccg atcgttccgt tcctgctgaa
     1381 cctgttcctg ggttttggca ttggtagctt tgcgcaaggc gacatcctgg gtggcagcct
     1441 gattctgggt ttcgatgcgg tgggcatcgg tctgattctg gcgggcgcgt acctggacat
     1501 caaggcgctg gatggtatta ccaaaaaggc ggcgtttcag tggacctggg gcaaaggtgt
     1561 gatgctggcg ggtgtggtta cgatggcggt tacccgtctg accgagatta tcctgccgtt
     1621 cacctttgcg aacagctata accgtaaact gaagaacagc ctgaacgtgg cgctgggtgg
     1681 cttcgaaccg agctttgacg ttgcgatggg tcagagcagc gcgctgggct tcgagctgag
     1741 ctttaaaaag agctacatgc tgatcaaaaa gatttgcttc tgcctgctgc tgctgcaggc
     1801 gaagattatc ctgagccaag atcagcaaaa cctggaattt aacaacgagt acttctactt
     1861 caacttccaa agcgcgtact atccgccgca cgaactgggc accaacggca gcggtttcag
     1921 ccagagcttt aaacacccga acttccaaaa catcgacccg agcctgaaaa ttccggtgaa
     1981 ctactggggt ggcatcaagc tgattagcta cctgggttac tacaagaact tcaaggcgct
     2041 gaacaacccg aacagcatct tctttaagaa caacggcatc gacattgatc tgaacatcgg
     2101 tctgagcccg gtgattgtta gcttcaaaag caccgttagc ttcaccccga tcgcgtttct
     2161 gaacctgtac gcgagcaacg aactgggcat tggttggcag gcgtttggct tcaaaggcgt
     2221 gggtgttcac atcgagaacg gcaagtacag caacgcggtg gaattttata gcgagatcac
     2281 cgcgggtggc cgttttcagt tcgacctgaa cgcgattttc caaggcgagt ggacccacat
     2341 tatcaccgtt atcggtaacg actttaccta cctgattaac ccgcacgcga aaaaggatca
     2401 actgtggaaa tataaggcgg acgatggcaa gaacatcaac ggtatgctga ttaaaccgta
     2461 cgcgctgctg gcgtataaga tgccgatccc gctgaacacc attggcctgc tgtatgaagg
     2521 taaaacccag atcggcaagg agcgtaacat tagcagctgg aaaaacaagg gctggggtag
     2581 cgacatcttc taccacaaca tcagcattgt tgcgaacttt gagattatca aaccgctgac
     2641 cctgggcctg caattcaagc tgagcaccaa cccgacctat accagcaaca ccgcgggtct
     2701 ggcggacatc agcaagcgta ttgcgaccgg caacagctac ttttactatg atagcatcgg
     2761 tatgagcatc acctacaagt acatgaacaa gaagcacacc aacttcagcg tgctgctgct
     2821 gctgatcttt ctgctgattc tgagcttcgg tggctttggt tactatatct accagagcaa
     2881 actgaacgac aagaaccgtg aaatcatgct gaacgaggtg aagaacagcg ttattgatcg
     2941 taactacaaa aaggcgtata gcgttgcgaa actgctgcag gacaagtacc cgcaaaacga
     3001 ggacatcgcg atgctgacca acaccctggc ggaaattgcg aacagcagcc cgttcgagag
     3061 caaagacctg cagcgtgata gcgcgaacca aatcctggac aaaattaagg gccaggataa
     3121 caccaagacc aacgtgaacg aaaacttcga catcgcgttt aacaaccgtt acatcaaaga
     3181 cagcaccatt accgagaact atagcgatcg taacgacgat gttggtatcg aggacgaaga
     3241 tattagcgag ttcaaaaaga gcaaaatccc ggagaaaatt aagccgaaca ccaacccgaa
     3301 ggaagaggac cagattatcc aaagcccgaa cccgaaactg agcgtgaacg atcagaagaa
     3361 cctgtttaac ctggagaaac tgaaaaagaa cctgagcggt aaaagcaaca gcgaaaacat
     3421 cctgaacgac agccagaaaa ttgagaacga taagcaaaac accaacctga gcaaagaaaa
     3481 gaacagcgag aacatcctga aaaccccgga caacagcaag tacagcaaca acaacaacac
     3541 caccagcctg aaaaagatta gcagcaacag ccagaaggag agcgaactga gcccgccgag
     3601 ccaaaccatt atcggcaaaa tctaccgtcc gtacagctat ctgattaaaa aggaactgta
     3661 tgagatcctg gacgatatta acaccggtcg tgtgaccctg ggcaaaaacc gtctgaagga
     3721 gctgatcaaa aagggtctga gcaacaaatt ccagaaggtt aacgaactga ttgagaacag
     3781 caaaaacaag gaagcgagca acctgctgct gaccctgatc aaaaaggaca ttgagccgaa
     3841 cctgatcaac attccgaaag atccgtacaa aaaggaaatc ttccagctgg acaaagagga
     3901 caaaaagccg caatatctgg aagacctgaa aagcaaggtg cacagcatca agccgattga
     3961 tctggaaaac accaagagcc gtcagcaagc gatcaaagac ctgaacgagt ttctgaagaa
     4021 caacccgaac gatgcgcagg cgagcaaaac cctggcgcag gcgaacaaga ttcaacacct
     4081 ggaggacctg aaaagcaagg ttcacagcat caaaccgatc gatctggaaa ataccaaaag
     4141 ccgtcagcag gcgattaagg acctgaacga gttcctgaaa aataatccga acgatgcgca
     4201 agcgagcaag accctggcgc aagcgaataa aattcaacac ctggaagatc tgaagagcaa
     4261 agttcatagc attaaaccga ttgatctgga aaataccaag agccgtcagc aggcgatcaa
     4321 ggatctgaat gagtttctga aaaataaccc gaatgacgcg caggcgagca agactttagc
     4381 gcaagcgaat aagatccagc acctggagga tctgaagagc aaggtgcata gcattaagcc
     4441 gatcgatctg gaaaacacca aaagccgtca gcaagcgatt aaagatctga atgagttcct
     4501 gaagaataat ccgaatgacg cgcaagcgag caaaacttta gcgcaggcga acaaaatcca
     4561 acacctggaa gacctgaaga gcaaagttca cagcattaaa cctatcgatc tggaaaacac
     4621 taagagccgt cagcaagcta ttaaggatct gaacgagttt ctgaaaaaca atccgaatga
     4681 tgcgcaggcg agcaagacat tagcgcaggc gaacaagatc caacacctgg aggatctgaa
     4741 aagcaaagtg cacagcatca aacctataga tctggaaaat actaagagcc gtcagcaggc
     4801 tatcaaggat ctgaacgagt tcctgaagaa taacccgaat gatgcgcaag cgagcaaaac
     4861 attagctcaa gctaataaga ttcagcacct ggaagatctg aaaagcaagg ttcatagcat
     4921 taagcctatc gatctggaaa atactaaaag ccgtcagcaa gctataaagg atttaaacga
     4981 gttcctgaaa aacaatccga acgacgctca ggcgagcaaa actctggcgc aagcgtacga
     5041 aaacaacggc gacctgctga aagcggaaaa cgcgtatgag aaaattatca agctgaccaa
     5101 cacccaggaa gaccactaca aactgggtat tatccgtttc aagctgaaaa agtatgaaca
     5161 cagcatcgag agctttgacc aaaccattaa gctggacccg aagcacaaaa aggcgctgca
     5221 caacaaaggt atcgcgctga tgatgctgaa caagaacaaa aaggcgattg agagcttcga
     5281 gaaagcgatc cagattgaca agaactacgg caccgcgtac tatcaaaaag gcatcgcgga
     5341 ggaaaagaac ggtgacatgc agcaagcgtt cgcgagcttt aaaaacgcgt acaacctgga
     5401 taagaacccg aactatgcgc tgaaagcggg catcgttagc aacaacctgg gtaacttcaa
     5461 gcagagcgag gaatacctga acttctttaa cgcgaacgcg aaaaagccga acgaaatcgc
     5521 gatttataac ctgagcatcg cgaaatttga aaacaacaag ctggaggaaa gcctggagac
     5581 catcaacaaa gcgattgacc tgaacccgga aaagagcgag tacctgtatc tgaaagcgag
     5641 catcaacctg aaaaaggaga actaccagaa cgcgatcagc ctgtatagcc tggtgattga
     5701 aaagaacccg gagaacacca gcgcgtacat caacctggcg aaagcgtatg aaaagagcgg
     5761 taacaaaagc caggcgatta gcaccctgga gaaaattatc aacaaaaaca acaagctggc
     5821 gctgaacaac ctgggtatcc tgtacaaaaa ggagaagaac tatcaaaaag cgatcgaaat
     5881 tttcgagaaa gcgattatca acagcgacat tgaagcgaag tacaacctgg cgaccaccct
     5941 gatcgagatt aacgacaaca cccgtgcgaa ggatctgctg cgtgaataca ccaaactgaa
     6001 accgaacaac ccggaggcgc tgcatgcgct gggcattatc gaatataacg agaataacaa
     6061 cgatcagacc ctgcgtgaac tgatcaaaaa gttcccgaac tacaaaaaga acgagaacat
     6121 caaaaagatt atcggtatta agcttttaat taagtttaaa cgagtattct atagtctcac
     6181 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6241 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6301 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6361 tcgtgccagc tgcattaatg aatcggccaa cgcgaacccc ttgcggccgc ccgggccgtc
     6421 gaccaattct catgtttgac agcttatcat cgaatttctg ccattcatcc gcttattatc
     6481 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta
     6541 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     6601 gaagccatca caaacggcat gatgaacctg aatagccagc ggcatcagca ccttgtcgcc
     6661 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     6721 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     6781 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     6841 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     6901 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     6961 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
     7021 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     7081 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     7141 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     7201 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     7261 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     7321 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     7381 atcttccgtc acaggtattt attcgcgata agctcatgga gcggcgtaac cgtcgcacag
     7441 gaaggacaga gaaagcgcgg atctgggaag tgacggacag aacggtcagg acctggattg
     7501 gggaggcggt tgccgccgct gctgctgacg gtgtgacgtt ctctgttccg gtcacaccac
     7561 atacgttccg ccattcctat gcgatgcaca tgctgtatgc cggtataccg ctgaaagttc
     7621 tgcaaagcct gatgggacat aagtccatca gttcaacgga agtctacacg aaggtttttg
     7681 cgctggatgt ggctgcccgg caccgggtgc agtttgcgat gccggagtct gatgcggttg
     7741 cgatgctgaa acaattatcc tgagaataaa tgccttggcc tttatatgga aatgtggaac
     7801 tgagtggata tgctgttttt gtctgttaaa cagagaagct ggctgttatc cactgagaag
     7861 cgaacgaaac agtcgggaaa atctcccatt atcgtagaga tccgcattat taatctcagg
     7921 agcctgtgta gcgtttatag gaagtagtgt tctgtcatga tgcctgcaag cggtaacgaa
     7981 aacgatttga atatgccttc aggaacaata gaaatcttcg tgcggtgtta cgttgaagtg
     8041 gagcggatta tgtcagcaat ggacagaaca acctaatgaa cacagaacca tgatgtggtc
     8101 tgtcctttta cagccagtag tgctcgccgc agtcgagcga cagggcgaag ccctcggctg
     8161 gttgccctcg ccgctgggct ggcggccgtc tatggccctg caaacgcgcc agaaacgccg
     8221 tcgaagccgt gtgcgagaca ccgcggccgg ccgccggcgt tgtggatacc tcgcggaaaa
     8281 cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg actcacccgg
     8341 cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga gctggccagc
     8401 ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga tgtggacaag
     8461 cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg acagatgagg
     8521 ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc acctattgac
     8581 atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc gcccgttttt
     8641 cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa accttgtttt
     8701 taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc
     8761 tcgaaccctc ccggtcgagt gagcgaggaa gcaccaggga acagcactta tatattctgc
     8821 ttacacacga tgcctgaaaa aacttccctt ggggttatcc acttatccac ggggatattt
     8881 ttataattat tttttttata gtttttagat cttctttttt agagcgcctt gtaggccttt
     8941 atccatgctg gttctagaga aggtgttgtg acaaattgcc ctttcagtgt gacaaatcac
     9001 cctcaaatga cagtcctgtc tgtgacaaat tgcccttaac cctgtgacaa attgccctca
     9061 gaagaagctg ttttttcaca aagttatccc tgcttattga ctctttttta tttagtgtga
     9121 caatctaaaa acttgtcaca cttcacatgg atctgtcatg gcggaaacag cggttatcaa
     9181 tcacaagaaa cgtaaaaata gcccgcgaat cgtccagtca aacgacctca ctgaggcggc
     9241 atatagtctc tcccgggatc aaaaacgtat gctgtatctg ttcgttgacc agatcagaaa
     9301 atctgatggc accctacagg aacatgacgg tatctgcgag atccatgttg ctaaatatgc
     9361 tgaaatattc ggattgacct ctgcggaagc cagtaaggat atacggcagg cattgaagag
     9421 tttcgcgggg aaggaagtgg ttttttatcg ccctgaagag gatgccggcg atgaaaaagg
     9481 ctatgaatct tttccttggt ttatcaaacg tgcgcacagt ccatccagag ggctttacag
     9541 tgtacatatc aacccatatc tcattccctt ctttatcggg ttacagaacc ggtttacgca
     9601 gtttcggctt agtgaaacaa aagaaatcac caatccgtat gccatgcgtt tatacgaatc
     9661 cctgtgtcag tatcgtaagc cggatggctc aggcatcgtc tctctgaaaa tcgactggat
     9721 catagagcgt taccagctgc ctcaaagtta ccagcgtatg cctgacttcc gccgccgctt
     9781 cctgcaggtc tgtgttaatg agatcaacag cagaactcca atgcgcctct catacattga
     9841 gaaaaagaaa ggccgccaga cgactcatat cgtattttcc ttccgcgata tcacttccat
     9901 gacgacagga tagtctgagg gttatctgtc acagatttga gggtggttcg tcacatttgt
     9961 tctgacctac tgagggtaat ttgtcacagt tttgctgttt ccttcagcct gcatggattt
    10021 tctcatactt tttgaactgt aatttttaag gaagccaaat ttgagggcag tttgtcacag
    10081 ttgatttcct tctctttccc ttcgtcatgt gacttgatat cgggggttag ttcgtcatca
    10141 ttgatgaggg ttgattatca cagtttatta ctctgaattg gctatccgcg tgtgtacctc
    10201 tacctggagt ttttcccacg gtggatattt cttcttgcgc tgagcgtaag agctatctga
    10261 cagaacagtt cttctttgct tcctcgccag ttcgctcgct atgctcggtt acacggctgc
    10321 ggcg
//
