LOCUS       sequence-223900-        6630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral system for reduced ectopic expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  AUTHORS   Dean KM, Roudot P, Reis CR, Welf ES, Mettlen M, Fiolka R
  TITLE     Diagonally Scanned Light-Sheet Microscopy for Fast Volumetric 
            Imaging of Adherent Cells.
  JOURNAL   Biophys J. 2016 Mar 29;110(6):1456-65. doi: 
            10.1016/j.bpj.2016.01.029.
  PUBMED    27028654
REFERENCE   2  (bases 1 to 6630)
  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.bpj.2016.01"; journalName: "Biophys J"; date: 
            "2016-03-29- 29"; volume: "110"; issue: "6"; pages: "1456-65"
COMMENT     Sequence Label: pLVX-CMV200
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     misc_feature    2402..2458
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    2599..3187
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2652..2672)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3070..3081)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3394..4027
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4155..4171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4155..4171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4168..4190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4179..4195
                     /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(4203..4209,4210..4227,4228..4233))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4248..4269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4386..4403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4557..5145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4637..4656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5316..6107,6108..6176))
                     /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     5939..5958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6177..6281)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6329..6463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6366..6385)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6420..6439
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatagttat taatagtaat caattacggg gtcattagtt
     2221 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2281 ccgcccaacg acccccgccc attgacgtca ataatgacgt acgttcccat agtaacgcca
     2341 atagggactt tccattgacg tcaatgggtg gagtatttac ggtccgctag cgctaactag
     2401 tagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     2461 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2521 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2581 taacgacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2641 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2701 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2761 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2821 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2881 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2941 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     3001 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3061 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3121 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3181 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
     3241 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     3301 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     3361 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
     3421 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     3481 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     3541 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     3601 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     3661 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     3721 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     3781 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     3841 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     3901 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     3961 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4021 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     4081 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
     4141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6301 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
     6361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6421 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
     6481 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
     6541 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
     6601 ttgttaaata tgtactacaa acttagtagt
//
