LOCUS       sequence-288706-        6649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of MBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6649)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6649)
  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: pCIneo-lambdaN-HA-MBP_F
FEATURES             Location/Qualifiers
     source          1..6649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1185..1211
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2169..2178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2353..2474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2364..2383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2418..2437
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2660..3115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2747..2766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2957..2978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3232..3589
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3232..3423
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3255..3275)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3440..3575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3640..4434
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3694..3713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4304..4323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4498..4546
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4587..4606)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4706..4728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4766..4784)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4852..4956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4957..5025,5026..5817)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5175..5194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5988..6576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6477..6496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg agaattcaat ggcttaccca tacgatgttc
     1201 cagattacgc tatgaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag
     1261 gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca
     1321 ccgttgagca tccggataaa ctggaagaga aattcccgca ggttgcggca actggcgatg
     1381 gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt
     1441 tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg
     1501 ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga
     1561 tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg
     1621 ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact
     1681 tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt
     1741 acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg
     1801 ttgacctgat taaaagcaaa cacatgaatg cagacaccga ttactccatc gcagaagctg
     1861 cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg
     1921 acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca
     1981 aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg
     2041 cgaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag
     2101 acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg aaagatccac
     2161 gtattgccgc caccatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga
     2221 tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga
     2281 ctgtcgatga agccctgaaa gattaggcgg ccgcttccct ttagtgaggg ttaatgcttc
     2341 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2401 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2461 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     2521 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc cgataaggat
     2581 cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2641 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2701 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2761 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2821 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2881 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2941 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3001 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3061 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     3121 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     3181 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3241 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3301 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3421 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3481 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3541 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc
     3601 ttctgacaca acagtctcga acttaaggct agagccacca tgattgaaca agatggattg
     3661 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3721 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3781 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3841 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3901 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3961 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4021 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4081 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4141 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4201 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4261 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4321 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4381 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4441 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     4501 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     4561 aggatccgcg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4621 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4681 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4741 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4801 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4861 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4921 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4981 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5041 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5101 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5161 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5281 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5341 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5461 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5521 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5581 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5641 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5821 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5881 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6061 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6301 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6601 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatct
//
