LOCUS       sequence-361445-        8618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of palmitoylated TagRFP-T for live imaging of plasma 
            membrane (protrusions, tunneing nanotubes), in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8618)
  AUTHORS   Haimovich G, Ecker CM, Dunagin MC, Eggan E, Raj A, Gerst JE, Singer 
            RH
  TITLE     Intercellular mRNA trafficking via membrane nanotube-like extensions
            in mammalian cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 14;114(46):E9873-E9882. doi: 
            10.1073/pnas.1706365114. Epub 2017 Oct 24.
  PUBMED    29078295
REFERENCE   2  (bases 1 to 8618)
  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"; doi: 
            "10.1073/pnas.1706365114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-11-14- 14"; volume: "114"; issue: "46"; pages: 
            "E9873-E9882"
COMMENT     Sequence Label: pHAGE-UBC-TagRFP-T-ps
FEATURES             Location/Qualifiers
     source          1..8618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(120..188,189..980)
                     /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(338..357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1151..1739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1640..1659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1893..1910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2027..2048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2063..2068,2069..2086,2087..2093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2101..2117
                     /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     2106..2128
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2125..2141
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2125..2141
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2160..2180
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2162..2180
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2281..2660
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2661..2864
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2815..2835
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2879..3059
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3106..3231
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3724..3957
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4142..4186
                     /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    4453..4570
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4627..5838
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5675..5694
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    6669..7257
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6722..6742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7140..7151)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7344..7577
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7649..7770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7686..7705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7740..7759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7810..7945
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7965..7984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7966..7984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7994..8011)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7994..8010)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8003..8025)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8152..8607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8239..8258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8449..8470
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
       61 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1801 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1861 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1921 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1981 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     2041 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     2101 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     2161 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttggcca ttgcatacgt
     2221 tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt
     2281 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2341 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2401 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2461 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2521 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2581 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2641 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     2701 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     2761 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     2821 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccggg
     2881 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     2941 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3001 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     3061 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     3121 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     3181 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     3241 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     3301 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     3361 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     3421 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     3481 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     3541 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     3601 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     3661 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     3721 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     3781 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     3841 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     3901 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     3961 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     4021 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     4081 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     4141 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     4201 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     4261 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     4321 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     4381 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
     4441 gtatcggtta acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     4501 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     4561 tcaaaatttt atcgatcacg agactagcct cgagaagctt gattaacccg tgtcggctcc
     4621 agatctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc ctcacggcga
     4681 gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc cggacgctca
     4741 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     4801 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     4861 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     4921 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     4981 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     5041 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     5101 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     5161 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     5221 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     5281 ggccttcgct aatgcgggaa agctcttatt cgggtgagat gggctggggc accatctggg
     5341 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     5401 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     5461 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     5521 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     5581 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     5641 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     5701 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     5761 gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt
     5821 ttggcttttt tgttagacga agcttgggct gcaggtcgag atccaccggt atgctgtgct
     5881 gtatgagaag aaccaaacag gttgaaaaga atgatgagga ccaaaagatc gtgagcaagg
     5941 gcgaggagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     6001 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     6061 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     6121 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta
     6181 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     6241 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     6301 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     6361 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg
     6421 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     6481 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     6541 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     6601 actgcgacct ccctagcaaa ctggggcaca aacttaatgg catggacgag ctgtacaagt
     6661 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     6721 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     6781 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     6841 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     6901 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6961 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     7021 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     7081 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     7141 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     7201 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     7261 gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     7321 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     7381 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7441 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7501 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7561 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     7621 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     7681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7741 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     7801 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     7861 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     7921 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttsgccctat agtgagtcgt
     7981 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     8041 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     8101 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     8161 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     8221 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     8281 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8341 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     8401 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     8461 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     8521 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     8581 ttaacaaaat attaacgctt acaatttagg tggcactt
//
