LOCUS       sequence-462722-        5054 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of His-tagged GNL2 (381-731aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5054)
  TITLE     Iuchi Lab GNL2 Plasmids 2025
REFERENCE   2  (bases 1 to 5054)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGNL2-#8
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1681..1698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1842..1963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1879..1898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1933..1952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1970..2425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2107..2128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2319..2338
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2452..2556
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2558..2915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2558..2749
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2581..2601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2766..2901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2950..3744
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2950..3744
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3004..3023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3614..3633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3932..3951)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3976..4023
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4352..4940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4841..4860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgaa gcccaaggtt gttcaagtag aaaaaattaa gagtcctgaa
      661 gaccacattg gtgctgtact tgaacgagca aagccagaat atatcagcaa aacatacaag
      721 attgattctt gggagaatgc tgaggacttt cttgagaagc tcgctttccg gactgggaag
      781 ttactaaagg gtggagagcc cgacttgcag actgtgggta agatggtcct caatgactgg
      841 cagaggggcc ggattccttt ctttgtcaag ccacccaatg cagagccact tgtggccccc
      901 cagcttctac cctcctcatc tttggaagtt gtcccagaag cagcccagaa caatccaggg
      961 gaggaagtca cagaaactgc aggtgaaggg tcagaatcca tcattaagga agaaacagaa
     1021 gagaacagtc actgtgatgc taacacagag atgcagcaga ttctcacacg agttcggcag
     1081 aactttggta aaatcaacgt ggtgcctcag ttttctgggg atgacctggt tcctgtggag
     1141 gtgtcagatc ttgaggaaga gcttgagagc ttttctgatg aagaggagga ggaacaggag
     1201 caacaaagag atgatgcgga agagtcttcc tcggagcctg aggaggaaaa tgtgggaaac
     1261 gacaccaaag ccgttattaa agcactggat gagaagattg ccaaatatca gaagtttcta
     1321 gacaaagcca aagccaaaaa gttttcagca gtcagaatat ccaagggact gagtgaaaag
     1381 atatttgcaa aacctgaaga acaaagaaaa acactggaag aagatgtaga tgacagagca
     1441 ccttccaaaa agggaaagaa gcggaaggca caaagggaag aggaacagga acattcaaat
     1501 aaagctccca gggcgcttac atcaaaagaa cggaggcgag cagtacgaca gcaacggccg
     1561 aaaaaagttg gtgtgcgcta ctatgaaaca cacaacgtga aaaataggaa caggaacaaa
     1621 aagaagacca atgactcaga gggacagaaa cacaaacgca aaaaattcag acaaaagcag
     1681 caccaccacc accaccactg accgcgggcc cgggatccac cggatctaga taactgatca
     1741 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1801 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1861 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1921 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     1981 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2041 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2101 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2161 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2221 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2281 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2341 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2401 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2461 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2521 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     2581 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2641 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2761 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2821 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2881 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     2941 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3001 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3061 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3121 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3181 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3241 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3301 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3361 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3421 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3481 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3541 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3601 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3661 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3721 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3781 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3841 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3901 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3961 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4021 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4081 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4141 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4981 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5041 ttaccgccat gcat
//
