LOCUS       sequence-7022-ex        4884 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 4884)
  AUTHORS   Colwill K, Wells CD, Elder K, Goudreault M, Hersi K, Kulkarni S, 
            Hardy WR, Pawson T, Morin GB
  TITLE     Modification of the Creator recombination system for proteomics 
            applications--improved expression by addition of splice sites.
  JOURNAL   BMC Biotechnol. 2006 Mar 6;6:13. doi: 10.1186/1472-6750-6-13.
  PUBMED    16519801
REFERENCE   2  (bases 1 to 4884)
  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: "BMC 
            Biotechnol."; date: "2006-03-6"; volume: "6:13. doi"; pages: " 
            10.1186/1472-6750-6-13"
COMMENT     Sequence Label: V37 pDNR MCS SA - Donor
FEATURES             Location/Qualifiers
     source          1..4884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(235..329)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(350..1009)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     924..943
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(1038..1071)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1086..1531
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1532..1618,1619..2953)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        3021..3125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3126..3194,3195..3986)
                     /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(3344..3363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4157..4745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4646..4665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4861..4878
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4862..4878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatggcgcg agggtttcct tgacaatatc
       61 atacttatcc tgtccctttt ttttccccag gggcgcgccg tcgacggtac cggacatatg
      121 cccgggaatt cctgcaggat ccgctcgaga agctttctag accattcgtt tggcgcgcgg
      181 gccttaatta accaggtaag ttcctaggag atcctggtca tgactagtgc ttggattctc
      241 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
      301 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
      361 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
      421 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
      481 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
      541 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
      601 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
      661 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
      721 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
      781 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
      841 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
      901 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
      961 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     1021 tagctcctga aagatccata acttcgtata gcatacatta tacgaagtta tgcggccgcg
     1081 acgtccacat atacctgccg ttcactatta tttagtgaaa tgagatatta tgatattttc
     1141 tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca gaaactataa aaaatacaga
     1201 gaatgaaaag aaacagatag attttttagt tctttaggcc cgtagtctgc aaatcctttt
     1261 atgattttct atcaaacaaa agaggaaaat agaccagttg caatccaaac gagagtctaa
     1321 tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact gataaagcag gcaagaccta
     1381 aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc ttacatattt taggtctttt
     1441 tttattgtgc gtaactaact tgccatcttc aaacaggagg gctggaagaa gcagaccgct
     1501 aacacagtac ataaaaaagg agacatgaac gatgaacatc aaaaagtttg caaaacaagc
     1561 aacagtatta acctttacta ccgcactgct ggcaggaggc gcaactcaag cgtttgcgaa
     1621 agaaacgaac caaaagccat ataaggaaac atacggcatt tcccatatta cacgccatga
     1681 tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat caagttcctg agttcgattc
     1741 gtccacaatt aaaaatatct cttctgcaaa aggcctggac gtttgggaca gctggccatt
     1801 acaaaacgct gacggcactg tcgcaaacta tcacggctac cacatcgtct ttgcattagc
     1861 cggagatcct aaaaatgcgg atgacacatc gatttacatg ttctatcaaa aagtcggcga
     1921 aacttctatt gacagctgga aaaacgctgg ccgcgtcttt aaagacagcg acaaattcga
     1981 tgcaaatgat tctatcctaa aagaccaaac acaagaatgg tcaggttcag ccacatttac
     2041 atctgacgga aaaatccgtt tattctacac tgatttctcc ggtaaacatt acggcaaaca
     2101 aacactgaca actgcacaag ttaacgtatc agcatcagac agctctttga acatcaacgg
     2161 tgtagaggat tataaatcaa tctttgacgg tgacggaaaa acgtatcaaa atgtacagca
     2221 gttcatcgat gaaggcaact acagctcagg cgacaaccat acgctgagag atcctcacta
     2281 cgtagaagat aaaggccaca aatacttagt atttgaagca aacactggaa ctgaagatgg
     2341 ctaccaaggc gaagaatctt tatttaacaa agcatactat ggcaaaagca catcattctt
     2401 ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa cgcacggctg agttagcaaa
     2461 cggcgctctc ggtatgattg agctaaacga tgattacaca ctgaaaaaag tgatgaaacc
     2521 gctgattgca tctaacacag taacagatga aattgaacgc gcgaacgtct ttaaaatgaa
     2581 cggcaaatgg tacctgttca ctgactcccg cggatcaaaa atgacgattg acggcattac
     2641 gtctaacgat atttacatgc ttggttatgt ttctaattct ttaactggcc catacaagcc
     2701 gctgaacaaa actggccttg tgttaaaaat ggatcttgat cctaacgatg taacctttac
     2761 ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat gtcgtgatta caagctatat
     2821 gacaaacaga ggattctacg cagacaaaca atcaacgttt gcgcctagct tcctgctgaa
     2881 catcaaaggc aagaaaacat ctgttgtcaa agacagcatc cttgaacaag gacaattaac
     2941 agttaacaaa taaaaacgca aaagaaaatg ccgatatcct attggcattg acgtcaggtg
     3001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccttacgcg
     4861 tgtaaaacga cggccagtag atct
//
