LOCUS       sequence-6904-ex        7214 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 7214)
  TITLE     Shivdasani lab plasmids
REFERENCE   2  (bases 1 to 7214)
  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: p140 pETF-GST RanBP10 L301G
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             38..691
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             713..733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1970..1999
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2009..2032
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2095..2113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2109..2156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2193..2648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2280..2299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2490..2511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2741..3556)
                     /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(2741..3556)
                     /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     3464..3483
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3548..3567
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3678..4266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4167..4186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4420..4437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4452..4594
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4520..4539)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4680..4702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4696..4887)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5696..6778)
                     /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     6740..6759
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6779..6856)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7062..7081
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7169..7188
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7169..7187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7188..7212
                     /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)."
ORIGIN
        1 tctagattta aattaacttt aagaaggaga tatacatatg tcccctatac taggttattg
       61 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
      121 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
      181 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
      241 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
      301 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
      361 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
      421 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
      481 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
      541 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
      601 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
      661 cacgtttggt ggtggcgacc atcctccaaa agctagtctg gttccgagat ctgaaaacct
      721 gtattttcag ggatcgatcc caagcttgat ggcggcggcg acggcagacc cgggagctgg
      781 gaacccgcag gctggggact cctccggtgg ggactccggg ggcgggctac cgtcacctgg
      841 agagcaggaa ctgagtcggc gcctgcagcg cttgtaccct gcggttaatc agcacgagac
      901 gccgctgccg cgctcctgga gccccaagga caagtacaac tacatcggcc tctcccaggg
      961 aaacctccgt gtccattaca aaggtcatgg caaaaatcac aaagatgcag cctctgtacg
     1021 tgccacccac cccatcccag cagcctgtgg tatttactat tttgaagtaa agattgtcag
     1081 caaaggaaga gatgggtaca tgggaattgg actctcagcg caaggcgtta acatgaacag
     1141 actgcctggt tgggacaagc attcctatgg ttaccatggc gatgatgggc attccttctg
     1201 ctcctctggg acaggccagc cttatggtcc cacattcacc acaggggatg tgatcggctg
     1261 ttgtgtcaac ctcatcaatg gcacctgctt ctacaccaag aatggccaca gccttggtat
     1321 cgccttcaca gatctcccgg ccaacctcta ccccactgtc ggcctacaga cacctgggga
     1381 gattgtggat gccaactttg ggcagcagcc cttcctgttc gacattgagg actatatgcg
     1441 ggagtggcgt gccaaagtcc agggcactgt gcacggcttt cccataagtg cccggcttgg
     1501 cgagtggcag gcggtgctgc agaacatggt ctcgtcttac ctggtgcatc atgggtattg
     1561 ttccacagcc acagcttttg cccgaatgac tgaaaccccg attcaggaag agcaggcatc
     1621 cataaagaac agacaaaaaa tccagaaggg ggtgctggaa ggccgggtgg gtgaagccat
     1681 tgagacaacc cagcgcttct accctgggct gctggaacat aatcccaacc tgctcttcat
     1741 gctcaagtgt cgacagttcg tggagatggt gaacggcact gacagtgagg tccgcagcct
     1801 gagttcccga agccccaagt cccaggacag ctaccctggc tcccccagcc tcagcccccg
     1861 acatggcccc agtagttccc atatacacaa cacaggagca gacagtccca gctgcagcaa
     1921 tggcgtcgca tccaccaaga acaaacagaa ccacagtaaa taccccgcag aacaaaaact
     1981 catctcagaa gaggatctgt agctcgagca ccaccaccac caccaccacc actaattgat
     2041 taatacctag gctgctaaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2101 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2161 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2401 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2521 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2641 tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2701 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     2761 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2821 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2881 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2941 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3001 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3061 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     3121 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     3181 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     3241 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     3301 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     3361 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     3421 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     3481 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     3541 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     3601 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4021 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4081 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4141 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4201 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4261 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4321 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4381 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4441 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4501 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4561 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     4621 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4681 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     4741 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4801 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4861 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4921 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4981 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     5041 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     5101 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     5161 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     5221 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     5281 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     5341 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     5401 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     5461 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     5521 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     5581 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5641 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5761 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     5821 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     5881 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     5941 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     6001 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6061 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6121 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6181 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     6241 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     6301 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     6361 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     6421 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6481 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6541 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     6601 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     6661 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     6721 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     6781 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     6841 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     6901 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     6961 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     7021 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     7081 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     7141 atcgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     7201 gataacaatt cccc
//
