LOCUS       sequence-338528-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of no signal control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  TITLE     Rarebase PBC plasmids
REFERENCE   2  (bases 1 to 5354)
  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: EF1a-EGFP (no signal control)
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             52..846
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             52..846
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(106..125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     716..735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1034..1053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1078..1125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1454..2042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1943..1962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2206..3384
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2437..3375
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3332..3352
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3407..3416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3413..3415,3416..3418,3419..4129)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3413..3415,3416..3418,3419..4129)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3458..3479)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3719..3738)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4066..4087
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4298..4419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4335..4354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4389..4408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4426..4881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4563..4584)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4775..4794
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4908..5012
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5014..5354,1..17)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5014..5205
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5037..5057)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5222..5354,1..3)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
       61 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
      121 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
      181 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
      241 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
      301 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
      361 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
      421 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
      481 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
      541 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
      601 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
      661 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
      721 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
      781 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
      841 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
      901 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      961 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     1021 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     1081 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     1141 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     1201 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     1261 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     1321 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1381 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1561 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2101 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     2161 tattaatgct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
     2221 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     2281 tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     2341 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     2401 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
     2461 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
     2521 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
     2581 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     2641 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     2701 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     2761 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     2821 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     2881 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct ctcgccgccg
     2941 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     3001 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     3061 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     3121 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     3181 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     3241 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     3301 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     3361 tttcttccat ttcaggtgtc gtgacgtacg gctagcgcta ccggtcgcca ccatggtgag
     3421 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3481 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3541 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3601 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3661 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3721 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3781 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3841 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3901 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3961 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4021 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4081 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     4141 atctcgagct caagcttcga attcttaagg atccaccgga tctagataac tgatcataat
     4201 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4261 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     4441 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4501 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4561 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4621 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4681 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4741 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4801 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4861 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4921 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4981 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5041 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5101 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5161 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5221 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5281 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5341 aggctttttt ggag
//
