LOCUS       sequence-229633-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral expression under a CMVie promoter with 
            Blasticidin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Puleo JI, Parker SS, Roman MR, Watson AW, Eliato KR, Peng L, Saboda 
            K, Roe DJ, Ros R, Gertler FB, Mouneimne G
  TITLE     Mechanosensing during directed cell migration requires dynamic actin
            polymerization at focal adhesions.
  JOURNAL   J Cell Biol. 2019 Oct 8. pii: jcb.201902101. doi: 
            10.1083/jcb.201902101.
  PUBMED    31594807
REFERENCE   2  (bases 1 to 7235)
  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: "J Cell 
            Biol. 2019 Oct 8. pii: jcb.201902101. doi: 10.1083/jcb.201902101."
COMMENT     Sequence Label: pLenti CMVie-IRES-BlastR alt MCS
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2005..2208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2159..2179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2205..2229
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(2317..2891,2892..2894,2895..2903)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2484..2501)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2711..2730
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2910..3308
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    3331..3919
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3384..3404)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3802..3813)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3993..4226
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4298..4419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4335..4354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4389..4408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4438..4573
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4611..4627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4611..4627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4624..4646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4635..4651
                     /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(4659..4665,4666..4683,4684..4689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4704..4725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4842..4859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5772..6563,6564..6632))
                     /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     6395..6414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6633..6737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6805..6823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6861..6883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6983..7002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7196..7218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7210..7227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7211..7227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctaggac
     2281 cggtggatcc ctcgaggttt aaactctaga gatccgcccc tctccctccc ccccccctaa
     2341 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     2401 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     2461 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     2521 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     2581 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2641 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2701 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2761 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     2821 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     2881 acgatgataa gcttgccaca accaccggga tggccaagcc tttgtctcaa gaagaatcca
     2941 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     3001 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     3061 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     3121 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     3181 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     3241 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     3301 agggctaagc gcggccgcgg cggagtcgac aatcaacctc tggattacaa aatttgtgaa
     3361 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3421 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3481 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3541 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3601 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3661 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3721 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3781 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3841 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3901 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     3961 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     4021 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     4081 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4141 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4201 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     4261 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     4321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     4441 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4501 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4561 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6781 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6841 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6901 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6961 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     7021 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     7081 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7141 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7201 tcacgacgtt gtaaaacgac ggccagtgcc aagct
//
