LOCUS       sequence-46901-e        4829 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 4829)
  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 4829)
  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: V678 pDNR MCS SA ES - Donor
FEATURES             Location/Qualifiers
     source          1..4829
                     /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(176..270)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(291..950)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     865..884
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(979..1012)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1031..1476
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1477..1563,1564..2898)
                     /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        2966..3070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3071..3139,3140..3931)
                     /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(3289..3308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4102..4690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4591..4610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4806..4823
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4807..4823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatggcgcg agggtttcct tgacaatatc
       61 atacttatcc tgtccctttt ttttccccag gggcgcgaat tcagcggccg cgggatccgt
      121 cgacttaatt aaccaggtaa gttcctagga gatcctggtc atgactagtg cttggattct
      181 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
      241 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
      301 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
      361 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      421 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
      481 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
      541 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
      601 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
      661 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
      721 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
      781 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
      841 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
      901 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
      961 ttagctcctg aaagatccat aacttcgtat agcatacatt atacgaagtt atgcggccgg
     1021 ccgcgacgtc cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata
     1081 ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat
     1141 acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc
     1201 cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag
     1261 tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag
     1321 acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt
     1381 ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga
     1441 ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa
     1501 caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt
     1561 gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc
     1621 catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgagttc
     1681 gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg
     1741 ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca
     1801 ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc
     1861 ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa
     1921 ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca
     1981 tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc
     2041 aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc
     2101 aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta
     2161 cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct
     2221 cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa
     2281 gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca
     2341 ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta
     2401 gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg
     2461 aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa
     2521 atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc
     2581 attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac
     2641 aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc
     2701 tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc
     2761 tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc tagcttcctg
     2821 ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa
     2881 ttaacagtta acaaataaaa acgcaaaaga aaatgccgat atcctattgg cattgacgtc
     2941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3061 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3301 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3481 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3601 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3901 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3961 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4021 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4081 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4141 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4201 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4261 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4321 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4381 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4441 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4501 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4561 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4621 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4681 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4741 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4801 acgcgtgtaa aacgacggcc agtagatct
//
