LOCUS       sequence-446521-        8166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express in mammalian cells the mRFP1 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 8166)
  TITLE     Actin-Endosomes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8166)
  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            "Unpublished"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-mRFP1-Human UTROPHIN_1-261
FEATURES             Location/Qualifiers
     source          1..8166
                     /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             3591..4265
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3591..4265
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3721..3739)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3989..4008
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4183..4206
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    5067..5655
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5120..5140)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5538..5549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5742..5975
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6047..6168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6084..6103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6138..6157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6208..6343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6363..6382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6364..6382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6392..6409)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6392..6408)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6401..6423)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6550..7005
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6637..6656)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6847..6868
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7031..7135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7136..7204,7205..7996)
                     /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(7354..7373)
                     /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 aggatccacc atggcctcct
     3601 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     3661 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     3721 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
     3781 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     3841 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3901 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     3961 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     4021 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
     4081 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
     4141 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
     4201 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
     4261 gcgcctccgg aaccatggcc aagtatggag aacatgaagc cagtcctgac aatgggcaga
     4321 acgaattcag tgatatcatt aagtccagat ctgatgaaca caatgacgta cagaagaaaa
     4381 cctttaccaa atggataaat gctcgatttt caaagagtgg gaaaccaccc atcaatgata
     4441 tgttcacaga cctcaaagat ggaaggaagc tattggatct tctagaaggc ctcacaggaa
     4501 catcactgcc aaaggaacgt ggttccacaa gggtacatgc cttaaataac gtcaacagag
     4561 tgctgcaggt tttacatcag aacaatgtgg aattagtgaa tataggggga accgacattg
     4621 tggatggaaa tcacaaactg actttggggt tactttggag catcattttg cactggcagg
     4681 tgaaagatgt catgaaggat gtcatgtcgg acctgcagca gacgaacagt gagaagatcc
     4741 tgctcagctg ggtgcgtcag accaccaggc cctacagcca agtcaacgtc ctcaacttca
     4801 ccaccagctg gacagatgga ctcgccttta atgctgtcct ccaccgacat aaacctgatc
     4861 tcttcagctg ggataaagtt gtcaaaatgt caccaattga gagacttgaa catgccttca
     4921 gcaaggctca aacttatttg ggaattgaaa agctgttaga tcctgaagat gttgccgttc
     4981 ggcttcctga caagaaatcc ataattatgt atttaacatc tttgtttgag gtgctacctc
     5041 agcaagtcac catagactaa ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     5101 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5161 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5221 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5281 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5341 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5401 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5461 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5521 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5581 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5641 gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga cttacaaggc
     5701 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5761 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     5821 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     5881 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     5941 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6001 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6061 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6121 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6181 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6241 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     6301 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     6361 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     6421 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6481 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     6541 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6601 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6661 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6721 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6781 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6841 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6901 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6961 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     7021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     7081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     7141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     7201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     7261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     7321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     7441 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     7501 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     7561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7741 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7801 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7861 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7981 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     8041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8161 atcttc
//
