LOCUS       sequence-7021-ex        4827 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 4827)
  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 4827)
  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: V7 pDNR Dual - Donor
FEATURES             Location/Qualifiers
     source          1..4827
                     /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(178..272)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(293..952)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     867..886
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(981..1014)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1029..1474
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1475..1561,1562..2896)
                     /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        2964..3068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3069..3137,3138..3929)
                     /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(3287..3306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4100..4688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4589..4608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4804..4821
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4805..4821
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatggcgcg ccgtcgacgg taccggacat
       61 atgcccggga attcctgcag gatccgctcg agaagctttc tagaccattc gtttggcgcg
      121 cgggccttaa ttaaccaggt aagttcctag gagatcctgg tcatgactag tgcttggatt
      181 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
      241 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc
      301 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
      361 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
      421 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
      481 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
      541 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
      601 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
      661 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
      721 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
      781 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
      841 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
      901 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
      961 ccttagctcc tgaaagatcc ataacttcgt atagcataca ttatacgaag ttatgcggcc
     1021 gcgacgtcca catatacctg ccgttcacta ttatttagtg aaatgagata ttatgatatt
     1081 ttctgaattg tgattaaaaa ggcaacttta tgcccatgca acagaaacta taaaaaatac
     1141 agagaatgaa aagaaacaga tagatttttt agttctttag gcccgtagtc tgcaaatcct
     1201 tttatgattt tctatcaaac aaaagaggaa aatagaccag ttgcaatcca aacgagagtc
     1261 taatagaatg aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag caggcaagac
     1321 ctaaaatgtg taaagggcaa agtgtatact ttggcgtcac cccttacata ttttaggtct
     1381 ttttttattg tgcgtaacta acttgccatc ttcaaacagg agggctggaa gaagcagacc
     1441 gctaacacag tacataaaaa aggagacatg aacgatgaac atcaaaaagt ttgcaaaaca
     1501 agcaacagta ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc
     1561 gaaagaaacg aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca
     1621 tgatatgctg caaatccctg aacagcaaaa aaatgaaaaa tatcaagttc ctgagttcga
     1681 ttcgtccaca attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc
     1741 attacaaaac gctgacggca ctgtcgcaaa ctatcacggc taccacatcg tctttgcatt
     1801 agccggagat cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg
     1861 cgaaacttct attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt
     1921 cgatgcaaat gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt
     1981 tacatctgac ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa
     2041 acaaacactg acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa
     2101 cggtgtagag gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca
     2161 gcagttcatc gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca
     2221 ctacgtagaa gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga
     2281 tggctaccaa ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt
     2341 cttccgtcaa gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc
     2401 aaacggcgct ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa
     2461 accgctgatt gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat
     2521 gaacggcaaa tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat
     2581 tacgtctaac gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa
     2641 gccgctgaac aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt
     2701 tacttactca cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga ttacaagcta
     2761 tatgacaaac agaggattct acgcagacaa acaatcaacg tttgcgccta gcttcctgct
     2821 gaacatcaaa ggcaagaaaa catctgttgt caaagacagc atccttgaac aaggacaatt
     2881 aacagttaac aaataaaaac gcaaaagaaa atgccgatat cctattggca ttgacgtcag
     2941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3061 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3121 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3181 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3241 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3301 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3361 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3421 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3481 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3541 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3601 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3661 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3721 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3781 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3841 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3901 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3961 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4021 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4081 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4141 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4201 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4261 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4321 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4381 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4441 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4501 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4561 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4621 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4681 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4741 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccttac
     4801 gcgtgtaaaa cgacggccag tagatct
//
