LOCUS       sequence-333961-        5547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5547)
  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_C
FEATURES             Location/Qualifiers
     source          1..5547
                     /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"
     intron          889..1021
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1004..1023
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1066..1084
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1088..1097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1094..1159
                     /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"
     polyA_signal    1251..1372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1262..1281)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1316..1335
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1558..2013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1645..1664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1855..1876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2130..2487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2130..2321
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2153..2173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2338..2473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2538..3332
                     /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(2592..2611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3202..3221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3396..3444
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3485..3504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3604..3626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3664..3682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3750..3854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3855..3923,3924..4715)
                     /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(4073..4092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4886..5474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5375..5394
                     /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 gtttagtgac cgtcagatca ctagaagctt tattgcggta gtttatcaca
      781 gttaaattgc taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagtg
      841 actctcttaa ggtagccttg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      901 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      961 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     1021 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
     1081 taggctagcc accatggacg cacaaacacg acgacgtgag cgtcgcgctg agaaacaagc
     1141 tcaatggaaa gctgcaaacc caccgctcga gaattcacgc gtggtacctc tagagtcgac
     1201 ccgggcggcc gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat
     1261 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1321 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1381 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     1441 gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc
     1501 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     1561 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1621 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1681 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1741 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1801 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1861 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1921 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1981 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc
     2101 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     2161 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2401 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2461 ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac
     2521 ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2581 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2641 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2701 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2761 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2821 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2881 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2941 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3001 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3061 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3121 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3181 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3241 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3301 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3361 ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac
     3421 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta tggtgcactc
     3481 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3541 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3601 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3661 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3721 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3781 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3841 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3901 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3961 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4021 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4081 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4141 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4201 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4261 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4321 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4381 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4441 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4501 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4561 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4621 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4681 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4741 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4801 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4861 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4921 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4981 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5041 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5101 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5161 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5221 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5281 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5341 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5401 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5461 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5521 cttttgctca catggctcga cagatct
//
