LOCUS       sequence-213518-        4002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NTF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4002)
  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 4002)
  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: pET23b-NTF2
FEATURES             Location/Qualifiers
     source          1..4002
                     /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"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             503..520
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(573..591)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      587..634
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      671..1126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(758..777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     968..989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1152..1256
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1257..1325,1326..2117)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1475..1494)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2288..2876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2777..2796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3030..3047
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3062..3201
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3127..3146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3287..3309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3303..3494)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgggagaca agccaatttg ggagcagatt ggatccagct
      121 tcattcaaca ttactaccag ttatttgata atgatagaac ccaactaggc gcaatttaca
      181 ttgacgcgtc atgccttacg tgggaaggac aacagttcca ggggaaagct gccattgtgg
      241 agaagttgtc tagccttccg ttccagaaaa ttcagcacag catcaccgcg caggaccatc
      301 agcccactcc agatagctgc atcatcagca tggttgtggg ccagcttaag gcggatgaag
      361 accccatcat ggggttccac cagatgttcc tattaaagaa catcaacgat gcttgggttt
      421 gcaccaatga catgttcagg ctcgccctgc acaactttgg ctgaattcga gctccgtcga
      481 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
      541 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
      601 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
      661 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      721 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      781 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      841 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      901 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      961 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1021 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1081 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     1141 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1201 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1261 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1321 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1381 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1441 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1501 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1561 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1621 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1681 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1741 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1801 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1861 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1921 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1981 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2041 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2101 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2161 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2221 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2281 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2341 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2401 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2461 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2521 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2581 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2641 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2701 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2761 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2821 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2881 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2941 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3001 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3061 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     3121 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3181 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3241 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3301 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     3361 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     3421 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     3481 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     3541 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     3601 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     3661 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     3721 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     3781 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     3841 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     3901 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     3961 ggccaggacc caacgctgcc cgagatctcg atcccgcgaa at
//
