LOCUS       sequence-289521-        7133 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 7133)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7133)
  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-FLuc_O
FEATURES             Location/Qualifiers
     source          1..7133
                     /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      1093..1102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1099..2751
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1165..1183)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2837..2958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2848..2867)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2902..2921
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3144..3599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3231..3250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3441..3462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3716..4073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3716..3907
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3739..3759)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3924..4059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4124..4918
                     /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(4178..4197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4788..4807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4982..5030
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5071..5090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5190..5212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5250..5268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5336..5440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5441..5509,5510..6301)
                     /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(5659..5678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6472..7060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6961..6980
                     /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 taggctagca cagccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc
     1141 tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc
     1201 ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct
     1261 gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca
     1321 aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc
     1381 gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg
     1441 ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa
     1501 aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct
     1561 aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc
     1621 ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg
     1681 atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact
     1741 gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat
     1801 actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga
     1861 tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg
     1921 aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc
     1981 ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct
     2041 ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca
     2101 ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag
     2161 ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg
     2221 gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt
     2281 cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag
     2341 gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc
     2401 gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg
     2461 gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac
     2521 gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg
     2581 gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga
     2641 ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa
     2701 atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta actcgagaat
     2761 tcacgcgtgg tacctctaga gtcgacccgg gcggccgctt ccctttagtg agggttaatg
     2821 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2881 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2941 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3001 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa
     3061 ggatcgatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3121 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3301 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3481 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3541 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     3601 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     3661 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     3721 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3901 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3961 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4021 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     4081 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     4141 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4201 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4261 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4321 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4381 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4441 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4501 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4561 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4621 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4681 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4741 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4801 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4861 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4921 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     4981 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     5041 gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5101 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5221 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5281 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5341 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5401 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5461 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5581 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5641 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5821 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6001 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6061 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6121 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6301 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6361 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6421 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7081 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tct
//
