LOCUS       sequence-369189-        7651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Third generation lentiviral vector expressing Reticulon 2 disease 
            mutant R60f fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7651)
  AUTHORS   Zamponi E, Meehl JB, Voeltz GK
  TITLE     The ER ladder is a unique morphological feature of developing 
            mammalian axons.
  JOURNAL   Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 
            10.1016/j.devcel.2022.05.002. Epub 2022 May 23.
  PUBMED    35609616
REFERENCE   2  (bases 1 to 7651)
  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: 
            "10.1016/j.devcel.2022.05.002"; journalName: "Dev Cell"; date: 
            "2022-06-6- 6"; volume: "57"; issue: "11"; pages: "1369-1382.e6"
COMMENT     Sequence Label: pLV-Rtn2b-R60f-mCherry
FEATURES             Location/Qualifiers
     source          1..7651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             218..928
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(363..381)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     631..650
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    944..1532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(997..1017)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1415..1426)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1619..1852
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1924..2045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1961..1980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2015..2034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2085..2220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2240..2259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2241..2259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2269..2286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2269..2285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2278..2300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2427..2882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2514..2533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2724..2745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2908..3012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3013..3081,3082..3873)
                     /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(3231..3250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4044..4632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4533..4552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4786..4803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4920..4941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4956..4961,4962..4979,4980..4986)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4994..5010
                     /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     4999..5021
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5018..5034
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5018..5034
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5053..5073
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5055..5073
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5101..5327
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5328..5508
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5555..5680
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6173..6406
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6591..6635
                     /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    6902..7018
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        7041..7344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7345..7548
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7499..7519
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7545..7569
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gatggggcag gtcctgccgg tcttcgccca ctgcaaagaa gctccgtcta cagcctcctc
       61 aactcctgat tccacagaag gagggaacga cgactctgat tttcgagagc tgcacacagc
      121 ccgggaattc tcagaggagg acgaggagga gaccacgtcg caggactggg gcaccccccc
      181 gggagctgac cttctcctac atcgccttgc accggtcatg gtgagcaagg gcgaggagga
      241 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      301 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      361 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      421 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      481 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      541 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
      601 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
      661 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      721 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
      781 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
      841 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
      901 cggcggcatg gacgagctgt acaagtaaag cggccgcgtc gacaatcaac ctctggatta
      961 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1021 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1081 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     1141 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     1201 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     1261 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1321 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     1381 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     1441 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     1501 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt acctttaaga
     1561 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     1621 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     1681 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     1741 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     1801 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     1861 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     1921 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1981 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2041 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     2101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2221 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     2281 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2341 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2401 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     2461 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2521 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2581 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2641 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2701 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2761 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2821 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     2881 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3001 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3121 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3181 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3241 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3301 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3361 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3421 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3481 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3541 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3601 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3661 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3721 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3781 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3841 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3901 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3961 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4021 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4081 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4141 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4201 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4261 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4321 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4381 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4441 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4501 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4561 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4621 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4681 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4741 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4801 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4861 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4921 aatgtgagtt agctcactca ttaggcaccc caggctttcc actttatgct tccggctcgt
     4981 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5041 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa
     5101 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
     5161 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
     5221 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
     5281 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
     5341 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5401 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5461 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     5521 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     5581 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     5641 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     5701 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     5761 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     5821 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     5881 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     5941 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     6001 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     6061 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     6121 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     6181 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     6241 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     6301 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     6361 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     6421 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     6481 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     6541 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     6601 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     6661 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     6721 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     6781 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     6841 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     6901 cttttaaaag aaaagggggg attggggggt atagtgcagg ggaaagaata gtagacataa
     6961 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat
     7021 cgataagctt gggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     7081 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     7141 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     7201 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     7261 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     7321 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     7381 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     7441 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     7501 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     7561 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgactc
     7621 tagagctagc gctaccggac tcagatctcg a
//
