LOCUS       sequence-347061-        6017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing five copies of the MUC1 tandem repeat (MUC1-5TR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6017)
  AUTHORS   Brokx RD, Revers L, Zhang Q, Yang S, Mal TK, Ikura M, Gariepy J
  TITLE     Nuclear magnetic resonance-based dissection of a glycosyltransferase
            specificity for the mucin MUC1 tandem repeat.
  JOURNAL   Biochemistry. 2003 Dec 2;42(47):13817-25. doi: 10.1021/bi0353070.
  PUBMED    14636048
REFERENCE   2  (bases 1 to 6017)
  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"; doi: "10"; journalName: 
            "Biochemistry"; date: "2003-12-2- 2"; volume: "42"; issue: "47"; 
            pages: "13817-25"
COMMENT     Sequence Label: Mucin MUC1-5TR-pET15b
FEATURES             Location/Qualifiers
     source          1..6017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(648..665)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(675..692)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             711..733
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    748..772
                     /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)."
     primer_bind     complement(772..791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(773..791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(875..894)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1100..1177
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1178..2260
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1197..1216)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3572..3763
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3757..3779)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3865..4004
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3920..3939
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4949..5740,5741..5809))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5572..5591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5810..5914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5982..6000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctattac ccgggcgctg gtcgagtatc tggcgcagaa
      361 gtaacaccat gagctggagg cgcagtagaa cctggagcag gccgagtgtc aggcgcagaa
      421 gtaacgccat gcgcaggggg cgcagtagag ccgggagcag gtcgagtatc aggagcagaa
      481 gtaaccccat gcgcgggagg agcagtagaa ccaggagcag gacgagtatc gggagcagaa
      541 gtcacaccat gagcgggtgg agcagtagag ccaggcgcgg ggcgagtatc tggagcagaa
      601 gtcacgccat gcgctggtgg cgcagtagac ccgggcgctg gcatatggct gccgcgcggc
      661 accaggccgc tgctgtgatg atgatgatga tggctgctgc ccatggtata tctccttctt
      721 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
      781 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc
      841 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
      901 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
      961 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     1021 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     1081 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     1141 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     1201 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     1261 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     1321 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     1381 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     1441 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     1501 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     1561 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     1621 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     1681 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     1741 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     1801 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     1861 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     1921 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     1981 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     2041 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     2101 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     2161 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2221 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     2281 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
     2341 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2401 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     2461 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     2521 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     2581 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     2641 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     2701 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     2761 tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg
     2821 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     2881 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag
     2941 ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg
     3001 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
     3061 cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat
     3121 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     3181 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     3241 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     3301 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     3361 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     3421 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     3481 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     3541 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     3601 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     3661 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     3721 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3781 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3841 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3901 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt
     3961 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     5281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     5581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5941 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6001 cctttcgtct tcaagaa
//
