LOCUS       sequence-7028-ex        4777 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 4777)
  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 4777)
  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: V795 pDNR MCS ES - Donor
FEATURES             Location/Qualifiers
     source          1..4777
                     /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(124..218)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(239..898)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     813..832
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(927..960)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        979..1424
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(1425..1511,1512..2846)
                     /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        2914..3018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3019..3087,3088..3879)
                     /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(3237..3256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4050..4638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4539..4558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4754..4771
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4755..4771
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatggcgcg ccgggaattc agcggccgcg
       61 ggatccgtcg acttaattaa ccaggtaagt tcctaggaga tcctggtcat gactagtgct
      121 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
      181 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
      241 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
      301 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
      361 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
      421 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
      481 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
      541 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
      601 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
      661 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
      721 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
      781 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
      841 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
      901 tagcttcctt agctcctgaa agatccataa cttcgtatag catacattat acgaagttat
      961 gcggccggcc gcgacgtcca catatacctg ccgttcacta ttatttagtg aaatgagata
     1021 ttatgatatt ttctgaattg tgattaaaaa ggcaacttta tgcccatgca acagaaacta
     1081 taaaaaatac agagaatgaa aagaaacaga tagatttttt agttctttag gcccgtagtc
     1141 tgcaaatcct tttatgattt tctatcaaac aaaagaggaa aatagaccag ttgcaatcca
     1201 aacgagagtc taatagaatg aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag
     1261 caggcaagac ctaaaatgtg taaagggcaa agtgtatact ttggcgtcac cccttacata
     1321 ttttaggtct ttttttattg tgcgtaacta acttgccatc ttcaaacagg agggctggaa
     1381 gaagcagacc gctaacacag tacataaaaa aggagacatg aacgatgaac atcaaaaagt
     1441 ttgcaaaaca agcaacagta ttaaccttta ctaccgcact gctggcagga ggcgcaactc
     1501 aagcgtttgc gaaagaaacg aaccaaaagc catataagga aacatacggc atttcccata
     1561 ttacacgcca tgatatgctg caaatccctg aacagcaaaa aaatgaaaaa tatcaagttc
     1621 ctgagttcga ttcgtccaca attaaaaata tctcttctgc aaaaggcctg gacgtttggg
     1681 acagctggcc attacaaaac gctgacggca ctgtcgcaaa ctatcacggc taccacatcg
     1741 tctttgcatt agccggagat cctaaaaatg cggatgacac atcgatttac atgttctatc
     1801 aaaaagtcgg cgaaacttct attgacagct ggaaaaacgc tggccgcgtc tttaaagaca
     1861 gcgacaaatt cgatgcaaat gattctatcc taaaagacca aacacaagaa tggtcaggtt
     1921 cagccacatt tacatctgac ggaaaaatcc gtttattcta cactgatttc tccggtaaac
     1981 attacggcaa acaaacactg acaactgcac aagttaacgt atcagcatca gacagctctt
     2041 tgaacatcaa cggtgtagag gattataaat caatctttga cggtgacgga aaaacgtatc
     2101 aaaatgtaca gcagttcatc gatgaaggca actacagctc aggcgacaac catacgctga
     2161 gagatcctca ctacgtagaa gataaaggcc acaaatactt agtatttgaa gcaaacactg
     2221 gaactgaaga tggctaccaa ggcgaagaat ctttatttaa caaagcatac tatggcaaaa
     2281 gcacatcatt cttccgtcaa gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg
     2341 ctgagttagc aaacggcgct ctcggtatga ttgagctaaa cgatgattac acactgaaaa
     2401 aagtgatgaa accgctgatt gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg
     2461 tctttaaaat gaacggcaaa tggtacctgt tcactgactc ccgcggatca aaaatgacga
     2521 ttgacggcat tacgtctaac gatatttaca tgcttggtta tgtttctaat tctttaactg
     2581 gcccatacaa gccgctgaac aaaactggcc ttgtgttaaa aatggatctt gatcctaacg
     2641 atgtaacctt tacttactca cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga
     2701 ttacaagcta tatgacaaac agaggattct acgcagacaa acaatcaacg tttgcgccta
     2761 gcttcctgct gaacatcaaa ggcaagaaaa catctgttgt caaagacagc atccttgaac
     2821 aaggacaatt aacagttaac aaataaaaac gcaaaagaaa atgccgatat cctattggca
     2881 ttgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2941 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3001 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3061 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3121 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3181 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3241 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3301 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3361 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3421 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3481 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3541 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3601 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3661 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3721 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3781 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3841 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3901 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3961 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4681 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4741 accgccttac gcgtgtaaaa cgacggccag tagatct
//
