LOCUS       sequence-212610-        6297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human RAN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  AUTHORS   Cassany A, Gerace L
  TITLE     Reconstitution of nuclear import in permeabilized cells.
  JOURNAL   Methods Mol Biol. 2009;464:181-205. doi: 
            10.1007/978-1-60327-461-6_11.
  PUBMED    18951186
REFERENCE   2  (bases 1 to 6297)
  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"; journalName: "Methods Mol
            Biol."; date: "2009"; volume: "464:181-205. doi"; pages: " 
            10.1007/978-1-60327-461-6_11"
COMMENT     Sequence Label: pET11d-RAN
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(799..817)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      813..860
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1035..1040,1041..1057,1058..1063))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1090..1108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1176..1280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1281..1349,1350..2141)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1499..1518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2312..2900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2801..2820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3054..3071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3086..3225
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3151..3170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3311..3333
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3327..3518)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5873..5892
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5912..5989)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6195..6214
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg gctgcgcagg gagagcccca ggtccagttc
      121 aaacttgtat tggttggtga tggtggtact ggaaaaacga ccttcgtgaa acgtcatttg
      181 actggtgaat ttgagaagaa gtatgtagcc accttgggtg ttgaggttca tcccctagtg
      241 ttccacacca acagaggacc tattaagttc aatgtatggg acacagccgg ccaggagaaa
      301 ttcggtggac tgagagatgg ctattatatc caagcccagt gtgccatcat aatgtttgat
      361 gtaacatcga gagttactta caagaatgtg cctaactggc atagagatct ggtacgagtg
      421 tgtgaaaaca tccccattgt gttgtgtggc aacaaagtgg atattaagga caggaaagtg
      481 aaggcgaaat ccattgtctt ccaccgaaag aagaatcttc agtactacga catttctgcc
      541 aaaagtaact acaactttga aaagcccttc ctctggcttg ctaggaagct cattggagac
      601 cctaacttgg aatttgttgc catgcctgct ctcgccccac cagaagttgt catggaccca
      661 gctttggcag cacagtatga gcacgactta gaggttgctc agacaactgc tctcccggat
      721 gaggatgatg acctgtgaga atgaagctgg atccggctgc taacaaagcc cgaaaggaag
      781 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      841 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
      901 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
      961 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     1021 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     1081 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1141 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1201 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1261 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1321 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1381 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1441 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1501 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1561 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1621 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1681 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1741 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1801 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     1861 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1921 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1981 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2041 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2101 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2161 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2221 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2281 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2341 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2401 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2461 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2521 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2581 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2641 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2701 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2761 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2821 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2881 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2941 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3001 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3061 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3121 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3181 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3241 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3301 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3361 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3421 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3481 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3541 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3601 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3661 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3721 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     3781 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     3841 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     3901 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     3961 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     4021 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     4081 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4141 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4201 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4261 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     4321 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4381 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4441 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4501 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4561 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4621 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4681 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4741 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4801 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaactgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4981 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5041 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5101 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     5161 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5221 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5281 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5341 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5401 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5461 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5521 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5581 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5641 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5701 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5761 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5821 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5881 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5941 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     6001 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6061 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6121 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6181 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6241 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
