LOCUS       sequence-347060-        5672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing CEA-N domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5672)
  AUTHORS   Abdul-Wahid A, Cydzik M, Prodeus A, Alwash M, Stanojcic M, Thompson 
            M, Huang EH, Shively JE, Gray-Owen SD, Gariepy J
  TITLE     Induction of antigen-specific TH 9 immunity accompanied by mast cell
            activation blocks tumor cell engraftment.
  JOURNAL   Int J Cancer. 2016 Aug 15;139(4):841-53. doi: 10.1002/ijc.30121. 
            Epub 2016 Apr 15.
  PUBMED    27037842
REFERENCE   2  (bases 1 to 5672)
  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: "10.1002/ijc.30121";
            journalName: "Int J Cancer"; date: "2016-08-15- 15"; volume: "139"; 
            issue: "4"; pages: "841-53"
COMMENT     Sequence Label: CEA-N domain residues 1-132 (CEACAM5)-pET30b
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(581..598)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             609..631
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    646..670
                     /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(670..689)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(671..689)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(777..796)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1002..1079
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1080..2162
                     /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(1099..1118)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2971..3162
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3156..3178)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3264..3403
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3319..3338
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3418..3435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3589..4177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3669..3688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4288..4307)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4299..5114
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4299..5114
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4372..4391)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5206..5661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5343..5364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5555..5574
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagctaggtg aaggccacag
      181 catccttgtc ctccacgggt ttggagttgt tgctggagat ggagggcttg ggcagctccg
      241 ggtatacccg gaactggcca gttgcttctt cattcacaag atctgacttt atgacgtgta
      301 gggtgtagaa tcctgtgtca ttctggatga tgttctggat cagcagggat gcattggggt
      361 atattatctc tcgaccactg tatgcgggcc ctggggtagc ttgttgagtt cctattacat
      421 atcctataat ttgacggttg ccatccactc tttcaccttt gtaccagctg tagccaaaaa
      481 gatgctgggg cagattgtgg acaagtagaa gcacctcctt cccctctgcg acattgaacg
      541 gcgtggattc aatagtgagc ttttggaaat agaggttttc atggtgatgg tgatgatgca
      601 tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
      661 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tctcgatcct
      721 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
      781 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
      841 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
      901 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
      961 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     1021 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     1081 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     1141 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     1201 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     1261 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     1321 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     1381 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     1441 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     1501 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     1561 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     1621 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     1681 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     1741 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     1801 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     1861 ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa gacagctcat
     1921 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     1981 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     2041 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     2101 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2161 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
     2221 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     2281 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     2341 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     2401 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     2461 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc
     2521 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     2581 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     2641 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
     2701 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     2761 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
     2821 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
     2881 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
     2941 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
     3001 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     3061 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     3121 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3181 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3241 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     3301 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg
     3361 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     3421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3481 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3541 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3601 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3661 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3721 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3781 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3841 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3901 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3961 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4021 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4081 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4141 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
     4261 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4321 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     4381 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     4441 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4501 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4561 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     4621 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     4681 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     4741 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     4801 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     4861 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     4921 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     4981 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     5041 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     5101 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5161 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5221 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5281 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5341 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5401 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5461 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5521 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5581 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5641 atgcgccgct acagggcgcg tcccattcgc ca
//
