LOCUS       sequence-486696-        6615 bp    DNA     circular SYN 18-SEP-2025
DEFINITION  Strep-Flag-MYCMutMBIIIa-His protein expression in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  TITLE     Myc-related plasmids
REFERENCE   2  (bases 1 to 6615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: M54
FEATURES             Location/Qualifiers
     source          1..6615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..23
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(76..94)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      90..137
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      174..629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(261..280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     471..492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(721..1536)
                     /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(721..1536)
                     /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     1444..1463
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1528..1547
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1658..2246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2147..2166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2400..2417
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2432..2571
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2497..2516)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2657..2679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2673..2864)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3673..4755)
                     /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     4717..4736
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4756..4833)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5039..5058
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5146..5165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5146..5164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5165..5189
                     /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)."
     CDS             5217..5240
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5247..5270
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6234..6260
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6504..6533
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6594..6611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg
       61 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
      121 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
      181 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      241 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      301 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      361 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      421 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      481 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      541 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      601 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      661 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
      721 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
      781 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      841 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      901 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      961 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     1021 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     1081 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     1141 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     1201 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     1261 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     1321 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     1381 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     1441 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     1501 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     1561 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     1621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1801 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2281 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2341 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2401 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2461 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     2521 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     2581 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2641 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2701 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     2761 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     2821 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     2881 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     2941 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     3001 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     3061 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     3121 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     3181 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     3241 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     3301 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     3361 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     3421 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     3481 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     3541 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     3601 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     3661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     3781 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     3841 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     3901 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
     3961 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     4021 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     4081 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     4141 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     4201 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     4261 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     4321 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     4381 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     4441 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     4501 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     4561 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     4621 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     4681 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     4741 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     4801 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     4861 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     4921 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     4981 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5041 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     5101 tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta
     5161 taggggaatt gtgagcggat aacaattccc ctctagaaag gagatatacc atggcttggt
     5221 cccatccgca gttcgagaaa ggttctgatt acaaggatga cgatgacaaa agtgggatgc
     5281 ctcttaacgt ctcgttcaca aaccgcaatt acgacttaga ctatgattcg gtacagccgt
     5341 atttctattg tgatgaagaa gagaatttct accagcagca acagcagtcc gagcttcagc
     5401 cacctgcccc ctctgaggat atctggaaaa aattcgaact tcttccaacg cccccgctgt
     5461 cgccgtcgcg ccgctcgggg ttatgcagcc ctagttacgt agccgtgacg cctttcagtc
     5521 ttcgtggcga caacgatggg ggtggcggaa gtttttcaac tgctgatcag cttgagatgg
     5581 tgactgagtt acttggggga gacatggtaa accagtcctt catttgtgat ccagacgacg
     5641 aaacttttat taagaacatt attatccaag actgcatgtg gagtggcttt tccgctgcag
     5701 ctaaattggt gtcagagaaa cttgcctcgt atcaagcggc gcgcaaagat tcgggaagtc
     5761 caaacccagc acgtggccat tcagtatgct caaccagttc gctttatctt caggatcttt
     5821 ccgccgctgc atcagaatgc gccgatccgt ccgctgcgtc accgtatccc gcgaacgata
     5881 gctcttcccc gaagtcgtgt gcttcacaag actcgtcggc attctcgcca tcctcggatt
     5941 ctctgctgtc ttcgactgag tcgtcgcctc aggggtctcc agagccttta gtccttcatg
     6001 aagagacacc accaactaca tcatctgact ccgaggagga gcaagaagac gaagaggaaa
     6061 ttgacgtcgt gagtgtagag aaacgtcagg ctcccggcaa acgttcggaa tcaggctctc
     6121 cctcagccgg aggtcatagc aagccgcccc actcaccatt agtcttgaag cgttgtcacg
     6181 tgtcaactca ccaacataat tacgctgccc ccccgtccac ccgtaaggat tatccagcgg
     6241 caaagcgcgt gaagttagat tctgttcgcg tgctgcgtca gatcagtaat aatcgtaaat
     6301 gtacgtcccc acgttcatca gatacagagg aaaatgttaa gcgccgcaca cacaatgttc
     6361 ttgaacgcca acgccgtaat gaattgaaac gcagtttttt tgccttacgt gaccagattc
     6421 ccgagcttga gaataacgag aaggccccga aggttgtgat tttgaagaaa gcgacggcgt
     6481 acattctttc tgtccaagca gaagaacaaa agctgattag tgaagaagat ttattgcgta
     6541 agcgtcgtga acagcttaag cataagttgg agcaattacg taactcctgt gcgcatcatc
     6601 accaccatca ctgac
//
