LOCUS       sequence-451918-        8103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes siRNA resistant SigmaR1 protein labeled with mNeonGreen 
            (lentiviral vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8103)
  AUTHORS   Sawyer EM, Jensen LE, Meehl JB, Larsen KP, Petito DA, Hurley JH, 
            Voeltz GK
  TITLE     SigmaR1 shapes rough endoplasmic reticulum membrane sheets.
  JOURNAL   Dev Cell. 2024 Jul 2:S1534-5807(24)00382-4. doi: 
            10.1016/j.devcel.2024.06.005.
  PUBMED    38971154
REFERENCE   2  (bases 1 to 8103)
  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: "Dev Cell. 
            2024 Jul 2:S1534-5807(24)00382-4. doi: 
            10.1016/j.devcel.2024.06.005."
COMMENT     Sequence Label: siRNA-resistant pLV-SigmaR1-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..8103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    44..169
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             216..396
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(397..623)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(651..671)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(651..669)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(690..706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(690..706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(703..725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    714..730
                     /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)."
     promoter        complement(join(738..744,745..762,763..768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    783..804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(921..938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1092..1680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1172..1191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1851..2642,2643..2711))
                     /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     2474..2493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2712..2816)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2842..3297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2979..3000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3191..3210
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3424..3446
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3438..3455
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3439..3455
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3465..3484
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3465..3483
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3504..3639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3679..3800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3690..3709)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3744..3763
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3872..4105
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4192..4780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4298..4309
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4707..4727
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4787..5497)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      5233..5242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5494..5503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6258..6282)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6279..6482)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        6483..6786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    6809..6925
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7192..7236)
                     /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    7421..7654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gaattttttc ccatcgcgat ctaattctcc cccgcttaat actgacgctc tcgcacccat
       61 ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt actcaccagt cgccgcccct
      121 cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt
      181 tccctttcgc tttcaagtcc ctgttcgggc gccactgcta gagattttcc acactgacta
      241 aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac acactacttg
      301 aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag
      361 ctcccaggct cagatctggt ctaaccagag agacccgttt atgtatcgag ctaggcactt
      421 aaatacaata tctctgcaat gcggcaattc agtggttcgt ccaatccatg tcagacccgt
      481 ctgttgcctt cctaataagg cacgatcgta ccaccttact tccaccaatc ggcatgcacg
      541 gtgctttttc tctccttgta aggcatgttg ctaactcatc gttaccatgt tgcaagacta
      601 caagagtatt gcataagact acattaagct tgcagctcca gcttttgttc cctttagtga
      661 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      721 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtggaaagc ctggggtgcc
      781 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      841 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      901 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2821 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2881 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2941 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3001 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3061 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3121 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3181 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3241 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     3301 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     3361 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     3421 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     3481 agggcgaatt gggtacgtcc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     3541 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     3601 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggat
     3661 agctagagcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3721 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3781 taagctgcaa taaacaagtt cctctcactc tctgatattc atttctttgc aagttataaa
     3841 tactgaataa taagatgaca tgaactacta ctgctagaga ttttccacac tgactaaaag
     3901 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     3961 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     4021 caggctcaga tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt
     4081 cgttgggagt gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct
     4141 acagctgcct tgtaagtcat tggtcttaaa ggtaccgagc tcgaattcca ggcggggagg
     4201 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     4261 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     4321 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccat ggaaaggacg
     4381 tcagcttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     4441 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     4501 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     4561 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     4621 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     4681 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     4741 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttac ttgtacagct
     4801 cgtccatgcc catcacatcg gtaaaggcct tttgccactc cttgaagttg agctcggtct
     4861 tggagtgctt gagctccgtc ttacggaaca cgtacatcgg ctggttcttc agatagttag
     4921 ccgccattgg cttggcaaag gtgtaggtgg tccgcgcagt gctccggtag cgcttgccat
     4981 ttccagtggt gtaactccac ttaaaggtac tgatgatggt tttgtcgttg gggtaagtct
     5041 tcttcgacct gcaccagtcc gcagcggtca gcgagttggt catcacagga ccgtcagcag
     5101 ggaaaccagt ccccttcacc tgggcctctc ctttgatgtg gcttccctcg taggtgtagc
     5161 ggtagttaac agtaagggag gcaccatctt caaactgcat tgtgcgatgg acttggtagc
     5221 cggagccatc taccatggcg gcctggaaag gcgacatccc gtcagggtag ggcaggtact
     5281 gatggaagcc atacccgata tgagggacca gaatccaggg ggagaactgg aggtcaccct
     5341 tggtggactt caggtttaac tcctcataac catcatttgg attgccggtg ccctgaccca
     5401 ccatgtcaaa gtccacaccg ttgatggagc caaagatgtg taactcatgt gtcgctggga
     5461 gagaggccat gttatcctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg
     5521 cccgcggtac cctagggtcc tggccaaaga ggtatgtcgt caattccaag cggaggcccc
     5581 gagcatagga gcgaagagta tagaagaggg tgaggaagtc ctgggtgctg aagacagtgt
     5641 cggccagcgc gaaggccagg gtggatggga tgacgccccg gccgtactcg accatccaag
     5701 tattgggccc ccactccaca gctgttgcct caccaggccc gtgtactacc gtctcccctg
     5761 ggtagaagac ctcacttttg gtggtgccct ctctccactg gtggaaggtg ccagagatga
     5821 tggtatccga gatctcagcc cagtagcgcc ccgagtggcc gcgggagccc aaggcggtgc
     5881 cgaagagcag cacatactcg gacagcgagg cgtgcagaag gcacatggcg cccatccatc
     5941 caccagcgtt gacgaacacc cactgcagct cctcgtcggg cagcacgtgg cctgggtgca
     6001 gccgccgcag ctccacgatc agtctgctaa atgcgagttc gtggtccagc ccagcgtact
     6061 gccgcgccaa ctgcgctatc tcttcgcgct ggaagacgaa gctctgcgta cccagccaga
     6121 gccagacgac ctgggtcagc accgctgcga cagccaggag cagcgcggcc cacgcccacc
     6181 gccggcccac ggcccactgc attctagagt cggtgtcttc tatggaggtc aaaacagcgt
     6241 ggatggcgtc tccaggcgat ctgacggttc actaaacgag ctctgcttat atagacctcc
     6301 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     6361 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     6421 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     6481 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     6541 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     6601 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     6661 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     6721 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     6781 gtaacgcgga actcccaagc ttatcgataa aattttgaat tttgtaattt gtttttgtaa
     6841 ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg cactataccc
     6901 cccaatcccc ccttttcttt taaaagttaa ccgataccgt cgagatccgt tcactaatcg
     6961 aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg gcctgtcggg
     7021 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
     7081 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
     7141 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
     7201 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
     7261 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
     7321 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
     7381 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
     7441 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
     7501 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
     7561 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
     7621 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
     7681 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
     7741 atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag gtctgaagat
     7801 cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta
     7861 aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg
     7921 tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca
     7981 gtatttgtct acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta
     8041 gctccctgct tgcccatact atatgtttta atttatattt tttctttccc cctggcctta
     8101 acc
//
