LOCUS       sequence-413632-        5063 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Nucleocytoplasmic transport probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5063)
  TITLE     Roca-Cusachs Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5063)
  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: H_NES 41kDa (BFP)
FEATURES             Location/Qualifiers
     source          1..5063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        323..526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     477..497
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             580..606
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(622..624,625..1335)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPIQVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLE
                     FRTAAGITLGMDELYK"
     CDS             1704..1730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1743..1760
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1813..1831)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1827..1874
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1900..1919
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1937..2392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2074..2095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2286..2305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2419..2523
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2525..2882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2525..2716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2548..2568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2733..2868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2917..3711
                     /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             2917..3711
                     /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(2971..2990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3581..3600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3899..3918)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3943..3990
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4319..4907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4808..4827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
       61 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      241 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      481 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      541 tcagatccgc tagcgctacc ggtcgccacc atgcttcaac ttcctcctct tgagagactt
      601 actcttggag gcgggggcag cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      661 atccaggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc
      721 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      781 cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc
      841 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      901 caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
      961 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcgtcgactt caaggaggac
     1021 ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat ctatatcatg
     1081 gccgtcaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa cgtggaggac
     1141 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1201 ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga ccccaacgag
     1261 aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac tctcggcatg
     1321 gacgagctgt acaagaagct tgctcaacaa aatgcttttt atcaagtctt aaatatgcct
     1381 aacttaaatg ctgatcaacg caatggtttt atccaaagcc ttaaagatga tccaagccaa
     1441 agtgctaacg ttttaggtga agctcaaaaa cttaatgact ctcaagctcc aaaagctgat
     1501 gcgcaacaaa ataacttcaa caaagatcaa caaagcgcct tctatgaaat cttgaacatg
     1561 cctaacttaa acgaagcgca acgtaacggc ttcattcaaa gtcttaaaga cgacccaagc
     1621 caaagcacta acgttttagg tgaagctaaa aaattaaacg aatctcaagc accgaaagct
     1681 gataacaatt aagtcgaccc gggtatccgt atgatgtgcc tgactacgca tgatatctcg
     1741 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     1801 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1861 tgaggggttt tttgctgaaa ggaggaacta tatccggatg tggtttgtcc aaactcatca
     1921 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1981 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2041 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2101 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2161 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2221 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2281 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2341 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     2401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2521 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2641 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2701 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2821 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2881 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     2941 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3001 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3061 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3121 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3181 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3241 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3301 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3361 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3421 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     3481 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3541 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3601 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3661 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3721 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3781 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3841 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     3901 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3961 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4021 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4081 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4141 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4201 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4261 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4321 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4381 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4801 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4861 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4921 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4981 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
     5041 attacggggt cattagttca tag
//
