LOCUS       sequence-399874-        5916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucleocytoplasmic transport probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5916)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5916)
  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: Diffusive 80kDa
FEATURES             Location/Qualifiers
     source          1..5916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      554..563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(560..562,563..565,566..1276)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(560..562,563..565,566..1276)
                     /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(605..626)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(866..885)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1213..1234
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2727..2745)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2741..2788
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2814..2833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2851..3306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2988..3009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3200..3219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3333..3437
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3462..3482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3506..3716
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3528..3724
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3575..3710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3759..4553
                     /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             3759..4553
                     /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(3813..3832)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4423..4442
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4741..4760)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4785..4832
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5161..5749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5650..5669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      541 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      601 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      661 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      721 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      781 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      841 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      901 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      961 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1021 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1081 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1141 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1201 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1261 ggacgagctg tacaagaagc ttgctcaaca aaatgctttt tatcaagtct taaatatgcc
     1321 taacttaaat gctgatcaac gcaatggttt tatccaaagc cttaaagatg atccaagcca
     1381 aagtgctaac gttttaggtg aagctcaaaa acttaatgac tctcaagctc caaaagctga
     1441 tgcgcaacaa aataacttca acaaagatca acaaagcgcc ttctatgaaa tcttgaacat
     1501 gcctaactta aacgaagcgc aacgtaacgg cttcattcaa agtcttaaag acgacccaag
     1561 ccaaagcact aacgttttag gtgaagctaa aaaattaaac gaatctcaag caccgaaagc
     1621 tgataacaat ttcaacaaag aacaacaaaa tgctttctat gaaatcttga atatgcctaa
     1681 cttaaacgaa gaacaacgca atggtttcat ccaaagctta aaagatgacc caagccaaag
     1741 tgctaaccta ttgtcagaag ctaaaaagtt aaatgaatct caagcaccga aagcggataa
     1801 caaattcaac aaagaacaac aaaatgcttt ctatgaaatc ttacatttac ctaacttaaa
     1861 cgaagaacaa cgcaatggtt tcatccaaag cctaaaagat gacccaagcc aaagcgctaa
     1921 ccttttagca gaagctaaaa agctaaatga tgctcaagca ccaaaagctc aacaaaatgc
     1981 tttttatcaa gtcttaaata tgcctaactt aaatgctgat caacgcaatg gttttatcca
     2041 aagccttaaa gatgatccaa gccaaagtgc taacgtttta ggtgaagctc aaaaacttaa
     2101 tgactctcaa gctccaaaag ctgatgcgca acaaaataac ttcaacaaag atcaacaaag
     2161 cgccttctat gaaatcttga acatgcctaa cttaaacgaa gcgcaacgta acggcttcat
     2221 tcaaagtctt aaagacgacc caagccaaag cactaacgtt ttaggtgaag ctaaaaaatt
     2281 aaacgaatct caagcaccga aagctgataa caatttcaac aaagaacaac aaaatgcttt
     2341 ctatgaaatc ttgaatatgc ctaacttaaa cgaagaacaa cgcaatggtt tcatccaaag
     2401 cttaaaagat gacccaagcc aaagtgctaa cctattgtca gaagctaaaa agttaaatga
     2461 atctcaagca ccgaaagcgg ataacaaatt caacaaagaa caacaaaatg ctttctatga
     2521 aatcttacat ttacctaact taaacgaaga acaacgcaat ggtttcatcc aaagcctaaa
     2581 agatgaccca agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca
     2641 agcaccaaaa taactcgagc accaccacca ccactgagat ccggctgcta acaaagcccg
     2701 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2761 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatgtggttt
     2821 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     2881 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2941 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3001 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3061 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3121 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3181 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3241 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3301 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3361 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3421 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3721 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3781 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3841 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3901 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3961 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4021 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4081 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4141 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4201 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4261 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4321 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4381 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4441 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4501 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4561 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4621 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4681 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4741 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4801 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4861 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4921 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4981 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5281 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5821 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     5881 caattacggg gtcattagtt catagcccat atatgg
//
