LOCUS       sequence-464186-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of myosin-binding protein C, 
            cardiac-type in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5903)
  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"
COMMENT     Sequence Label: pAIP4_3CX2
FEATURES             Location/Qualifiers
     source          1..5903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     630..649
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     714..733
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      844..1432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1333..1352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1586..1603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1618..1757
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1683..1702)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1843..1865
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1859..2050)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2859..3941)
                     /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     3903..3922
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3942..4019)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4225..4244
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4332..4351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4332..4350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4351..4375
                     /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)."
     RBS             4390..4412
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4435..4464
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5095..5112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5165..5183)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5179..5226
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5263..5718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5350..5369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5560..5581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5810..5903,1..722))
                     /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             complement(join(5810..5903,1..722))
                     /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"
ORIGIN
        1 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
       61 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
      121 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
      181 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
      241 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
      301 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
      361 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
      421 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
      481 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
      541 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
      601 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
      661 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
      721 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
      781 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      841 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      901 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      961 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1021 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1081 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1141 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1201 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1261 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1321 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1381 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1441 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1501 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1561 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     1621 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     1681 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     1741 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     1801 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1861 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     1921 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     1981 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     2041 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     2101 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     2161 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     2221 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     2281 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     2341 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     2401 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     2461 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     2521 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     2581 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     2641 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     2701 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     2761 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     2821 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2941 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     3001 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     3061 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     3121 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     3181 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     3241 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     3301 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     3361 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     3421 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     3481 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     3541 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     3601 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     3661 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     3721 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     3781 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     3841 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     3901 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     3961 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     4021 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     4081 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     4141 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     4201 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     4261 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga
     4321 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     4381 agaaataatt ttgtttaact ttaagaagga gatatacata tgatggcgaa gattcatcac
     4441 catcatcacc atcaccatca ccacggtagc ctgcaagatt cggaagtcaa ccaagaagca
     4501 aagccggaag tcaagccgga agtgaaaccg gaaacccata ttaacctgaa agttagtgac
     4561 ggcagctctg aaattttctt taagatcaaa aagaccacgc cgctgcgtcg cctgatggaa
     4621 gcgtttgcca aacgtcaggg caaggaaatg gatagcctgc gtttcctgta tgacggtatt
     4681 cgcatccagg cagatcaagc gccggaagac ctggacatgg aagacaacga catcattgaa
     4741 gcccaccgtg aacagattgg cggctctgga ctatttgtaa tgaggcccca agatggagaa
     4801 gtcaccgtgg gcggttccat tacctttagc gcacgtgttg cgggtgcgct gctgaaaccg
     4861 ccggttgtca aatggtttaa gggcaaatgg gttgatttgt cgagcaaggt gggccaacat
     4921 ttacagctgc acgatagcta cgaccgtgct agcaaggtgt atctgttcga gctgcacatc
     4981 acggacgccc agccggcgtt tactggttct taccgctgcg aagtttccac caaggacaaa
     5041 ttcgattgta gcaatttcaa cttgaccgtg catgagggtt cgtaatgact cgagcaccac
     5101 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     5161 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     5221 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     5281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5401 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5701 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5761 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     5821 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     5881 gaaaaagccg tttctgtaat gaa
//
