LOCUS       sequence-337921-        6012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human Ran protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6012)
  TITLE     Zeng Lab Plasmids
REFERENCE   2  (bases 1 to 6012)
  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: pET28a-His6-Ran(Q69L)
FEATURES             Location/Qualifiers
     source          1..6012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             29..51
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             70..87
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             97..114
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             118..150
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             847..864
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(917..935)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      931..978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1015..1470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1102..1121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1312..1333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1562..2377)
                     /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(1562..2377)
                     /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     2285..2304
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2369..2388
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2499..3087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2988..3007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3241..3258
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3273..3412
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3338..3357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3498..3520
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3514..3705)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4514..5596)
                     /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     5558..5577
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5597..5674)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5880..5899
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5983..6002
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5983..6001
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6002..6012,1..14)
                     /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 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg
       61 ggcagcagcc atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg
      121 gctagcatga ctggtggaca gcaaatgggt cgcggatccg aattcgagct ccgtcgacgc
      181 atggctgcgc agggagagcc ccaggtccag ttcaaacttg tattggttgg tgatggtggt
      241 actggaaaaa cgaccttcgt gaaacgtcat ttgactggtg aatttgagaa gaagtatgta
      301 gccaccttgg gtgttgaggt tcatccccta gtgttccaca ccaacagagg acctattaag
      361 ttcaatgtat gggacacagc cggcctcgag aaattcggtg gactgagaga tggctattat
      421 atccaagccc agtgtgccat cataatgttt gatgtaacat cgagagttac ttacaagaat
      481 gtgcctaact ggcatagaga tctggtacga gtgtgtgaaa acatccccat tgtgttgtgt
      541 ggcaacaaag tggatattaa ggacaggaaa gtgaaggcga aatccattgt cttccaccga
      601 aagaagaatc ttcagtacta cgacatttct gccaaaagta actacaactt tgaaaagccc
      661 ttcctctggc ttgctaggaa gctcattgga gaccctaact tggaatttgt tgccatgcct
      721 gctctcgccc caccagaagt tgtcatggac ccagctttgg cagcacagta tgagcacgac
      781 ttagaggttg ctcagacaac tactctcccg gatgaggatg atgacctgtg agcggccgca
      841 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
      901 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      961 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     1021 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1081 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1141 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1201 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1261 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1321 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1381 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1441 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     1501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     1561 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1621 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1681 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1741 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1801 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1861 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1921 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     1981 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     2041 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     2101 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     2161 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     2221 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     2281 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2341 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     2401 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2641 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3121 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3181 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3241 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3301 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     3361 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     3421 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3481 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3541 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3601 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3661 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     3721 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     3781 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     3841 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     3901 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     3961 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     4021 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     4081 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     4141 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     4201 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     4261 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     4321 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     4381 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     4441 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     4501 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4621 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4681 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4741 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4801 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4861 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4921 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4981 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     5041 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     5101 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     5161 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     5221 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5281 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5341 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5401 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5461 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5521 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5581 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5641 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5701 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5761 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5821 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5881 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     5941 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
     6001 gggaattgtg ag
//
