LOCUS       sequence-288822-        5608 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5608)
  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: pCIneo-lambdaN-HA_G
FEATURES             Location/Qualifiers
     source          1..5608
                     /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      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1104..1169
                     /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             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1312..1433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1323..1342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1377..1396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1619..2074
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1706..1725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1916..1937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2191..2548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2191..2382
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2214..2234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2399..2534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2599..3393
                     /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(2653..2672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3263..3282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3457..3505
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3665..3687
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3725..3743)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3811..3915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3916..3984,3985..4776)
                     /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(4134..4153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4947..5535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5436..5455
                     /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 aattgcggcc accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agaattcacg cgtggtacct ctagagtcga
     1261 cccgggcggc cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca
     1321 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1381 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1441 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     1501 agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag
     1561 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     1621 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1681 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1741 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1801 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1861 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1921 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1981 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2041 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     2101 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     2161 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     2221 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2281 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2341 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2401 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2461 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2521 cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa
     2581 cttaaggcta gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2641 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2701 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2761 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     2821 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2881 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2941 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3001 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3061 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3121 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3181 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3241 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3301 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3361 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3421 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     3481 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact
     3541 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3601 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3661 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3721 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3781 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3841 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3901 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3961 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4021 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4081 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4141 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4201 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4261 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4321 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4381 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4441 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4501 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4561 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4621 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4681 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4741 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4801 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4861 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4921 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4981 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5041 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5101 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5161 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5221 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5281 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5341 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5401 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5461 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5521 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5581 ccttttgctc acatggctcg acagatct
//
