LOCUS       sequence-446518-        8210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express in mammalian cells the EGFP fused to residues 1-261 of Human
            Utrophin to detect filamentous actin in live, fix cels or tissues..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8210)
  TITLE     Actin-Endosomes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8210)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLV-EGFP-Human-UTROPHIN_1-261
FEATURES             Location/Qualifiers
     source          1..8210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2859..2975
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2998..3301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3302..3505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3456..3476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3502..3526
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3593..3595,3596..3598,3599..4309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3593..3595,3596..3598,3599..4309)
                     /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(3638..3659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3899..3918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4246..4267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5111..5699
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5164..5184)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5582..5593)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5786..6019
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6091..6212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6128..6147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6182..6201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6252..6387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6407..6426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6408..6426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6436..6453)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6436..6452)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6445..6467)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6594..7049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6681..6700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6891..6912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7075..7179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7180..7248,7249..8040)
                     /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(7398..7417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg agttccgcgt
     3001 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3061 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3181 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3241 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3301 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3361 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3421 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3481 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     3541 tccacgctgt tttgacctcc atagaagaca ccgactctag aggatccaaa ccatggtgag
     3601 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3661 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3721 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3781 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3841 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3901 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3961 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4021 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4081 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4141 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4201 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4261 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggaaccat
     4321 ggccaagtat ggagaacatg aagccagtcc tgacaatggg cagaacgaat tcagtgatat
     4381 cattaagtcc agatctgatg aacacaatga cgtacagaag aaaaccttta ccaaatggat
     4441 aaatgctcga ttttcaaaga gtgggaaacc acccatcaat gatatgttca cagacctcaa
     4501 agatggaagg aagctattgg atcttctaga aggcctcaca ggaacatcac tgccaaagga
     4561 acgtggttcc acaagggtac atgccttaaa taacgtcaac agagtgctgc aggttttaca
     4621 tcagaacaat gtggaattag tgaatatagg gggaaccgac attgtggatg gaaatcacaa
     4681 actgactttg gggttacttt ggagcatcat tttgcactgg caggtgaaag atgtcatgaa
     4741 ggatgtcatg tcggacctgc agcagacgaa cagtgagaag atcctgctca gctgggtgcg
     4801 tcagaccacc aggccctaca gccaagtcaa cgtcctcaac ttcaccacca gctggacaga
     4861 tggactcgcc tttaatgctg tcctccaccg acataaacct gatctcttca gctgggataa
     4921 agttgtcaaa atgtcaccaa ttgagagact tgaacatgcc ttcagcaagg ctcaaactta
     4981 tttgggaatt gaaaagctgt tagatcctga agatgttgcc gttcggcttc ctgacaagaa
     5041 atccataatt atgtatttaa catctttgtt tgaggtgcta cctcagcaag tcaccataga
     5101 ctaactcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5161 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5221 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5281 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5341 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5401 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5461 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5521 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5581 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5641 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5701 tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5761 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5821 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5881 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5941 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6001 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     6061 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     6121 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6181 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     6241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6301 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6361 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6421 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6481 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6541 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6661 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6721 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6781 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6901 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6961 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7021 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     7081 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7141 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7381 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7861 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8041 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8101 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8161 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
