LOCUS       sequence-369187-        7722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Third generation lentiviral vector expressing GFP-Sec61-beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7722)
  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 7722)
  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-GFP-Sec61-beta
FEATURES             Location/Qualifiers
     source          1..7722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      904..1492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1393..1412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1646..1663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1780..1801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1816..1821,1822..1839,1840..1846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1854..1870
                     /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     1859..1881
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1878..1894
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1878..1894
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1913..1933
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1915..1933
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1961..2187
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2188..2368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2415..2540
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3033..3266
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3451..3495
                     /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    3762..3878
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3901..4204
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4205..4408
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4359..4379
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4405..4429
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4497..5213
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             5229..5519
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     misc_feature    5526..6114
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5579..5599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5997..6008)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6201..6434
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6506..6627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6543..6562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6597..6616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6667..6802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6822..6841)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6823..6841)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6851..6868)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6851..6867)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6860..6882)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7009..7464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7096..7115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7306..7327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7490..7594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7595..7663,7664..7722,1..733)
                     /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"
ORIGIN
        1 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
       61 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      121 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
      181 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
      241 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
      301 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
      361 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
      421 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      481 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
      541 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
      601 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
      661 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      721 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
      781 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      841 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      901 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      961 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1021 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1081 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1141 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1201 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1261 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1321 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1381 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1441 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1501 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1561 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1621 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     1681 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1741 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1801 ttaggcaccc caggctttcc actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1861 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     1921 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     1981 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     2041 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     2101 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     2161 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     2221 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     2281 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     2341 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     2401 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     2461 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     2521 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     2581 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     2641 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     2701 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     2761 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     2821 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     2881 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     2941 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     3001 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     3061 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     3121 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     3181 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     3241 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     3301 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     3361 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     3421 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     3481 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     3541 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     3601 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     3661 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     3721 tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag aaaagggggg
     3781 attggggggt atagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     3841 aaagaattac aaaaacaaat tacaaaattc aaaattttat cgataagctt gggagttccg
     3901 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3961 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     4021 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     4081 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     4141 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     4201 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     4261 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     4321 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     4381 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     4441 ccatccacgc tgttttgacc tccatagaag acaccgactc tagaggatcc accggtatgg
     4501 tgagcaaggg cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg
     4561 atgtgaatgg ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca
     4621 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg
     4681 tgaccaccct gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc
     4741 acgacttctt caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg
     4801 aggatgacgg caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga
     4861 atcgcatcga gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga
     4921 tggagtacaa ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca
     4981 tcaaggtgaa cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc
     5041 actaccagca gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc
     5101 tgtccaccca gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact
     5161 tcggcttcgt gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac
     5221 tcagatctat gcctggtccg acccccagtg gcactaacgt gggatcctca gggcgctctc
     5281 ccagcaaagc agtggccgcc cgggcggcgg gatccactgt ccggcagagg aaaaatgcca
     5341 gctgtgggac aaggagtgca ggccgcacaa cctcggcagg caccgggggg atgtggcgat
     5401 tctacacaga agattcacct gggctcaaag ttggccctgt tccagtattg gttatgagtc
     5461 ttctgttcat cgcttctgta tttatgttgc acatttgggg caagtacact cgttcgtagg
     5521 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5581 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5641 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5701 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5761 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5821 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5881 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5941 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6001 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6061 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     6121 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6181 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     6241 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6301 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6361 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6421 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6481 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     6541 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6601 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     6661 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6721 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6781 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     6841 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6901 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6961 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     7021 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7081 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7141 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7201 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7261 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7321 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7381 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7441 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7501 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7561 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7621 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7681 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg gg
//
