LOCUS       sequence-290085-        7719 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 7719)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7719)
  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: pcDNA3.1-hFluc-V5_U
FEATURES             Location/Qualifiers
     source          1..7719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             901..2550
                     /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(967..985)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2551..2592
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        complement(3275..3293)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3276..3293)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3313..3330)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3319..3543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3589..4017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3676..3695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3886..3907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4026..4046)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4031..4360
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4273..4292
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4427..5221
                     /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(4481..4500)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5091..5110
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5395..5516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5432..5451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5486..5505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5565..5581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5565..5581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5578..5600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5589..5605
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5613..5619,5620..5637,5638..5643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5658..5679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5796..5813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5967..6552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6047..6066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6723..7514,7515..7583))
                     /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     7346..7365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7584..7688)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
      961 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     1021 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
     1081 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     1141 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     1201 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
     1261 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     1321 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
     1381 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     1441 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
     1501 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
     1561 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     1621 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     1681 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
     1741 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
     1801 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
     1861 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     1921 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     1981 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     2041 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     2101 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     2161 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     2221 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     2281 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     2341 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     2401 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     2461 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     2521 aaggccaaga agggcggaaa gatcgccgtg ggtaagccta tccctaaccc tctcctcggt
     2581 ctcgattcta cgtaagcggc cgccacagtc catgccatca ctgccaccca gaagactgtg
     2641 gatggcccct ccgggaaact gtggcgtgat ggccgcgggg ctctccagaa catcatccct
     2701 gcctctactg gcgctgccaa ggctgtgggc aaggtcatcc ctgagctgaa cgggaagctc
     2761 actggcatgg ccttccgtgt ccccactgcc aacgtgtcag tggtggacct gacctgccgt
     2821 ctagaaaaac ctgccaaata tgatgacatc aagaaggtgg tgaagcaggc gtcggagggc
     2881 cccctcaagg gcatcctggg ctacactgag caccaggtgg tctcctctga cttcaacagc
     2941 gacacccact cctccacctt cgacgctggg gctggcattg ccctcaacga ccactttgtc
     3001 aagctcattt cctggtatga caacgaattt ggctacagca acagggtggt ggacctcatg
     3061 gcccacatgg cctccaagga gtaagacccc tggacaacca gccccagcaa gagcacaaga
     3121 ggaagagaga gaccctcact gctggggagt ccctgccaca ctcagtcccc caccacactg
     3181 aatctcccct cctcacagtt tccatgtaga ccccttgaag aggggagggg cctagggagc
     3241 cgcaccttgt catgtaccag ctagagggcc ctattctata gtgtcaccta aatgctagag
     3301 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3361 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3421 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3481 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3541 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3601 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3661 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3721 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3781 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3841 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3901 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3961 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4021 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4381 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4441 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4501 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4561 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4621 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4681 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4741 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4801 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4861 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4921 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4981 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5041 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5101 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5161 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5221 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5281 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5341 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5401 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5521 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5581 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5641 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5761 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5821 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5881 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5941 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6001 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6061 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6121 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6181 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6241 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6301 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6361 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6421 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6481 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6841 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7021 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7201 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7681 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
