LOCUS       sequence-242009-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgITGB4-3 sequence: GAGGAGCGTAGGTCCTCGCAG. Lentiviral mouse U6 (mU6)
            promoter-driven expression vector that coexpessed Puro-T2A-mCherry 
            from a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8308)
  AUTHORS   Lee JY, Dominguez AA, Nam S, Stowers RS, Qi LS, Chaudhuri O
  TITLE     Identification of cell context-dependent YAP-associated proteins 
            reveals beta1 and beta4 integrin mediate YAP translocation 
            independently of cell spreading.
  JOURNAL   Sci Rep. 2019 Nov 20;9(1):17188. doi: 10.1038/s41598-019-53659-4.
  PUBMED    31748579
REFERENCE   2  (bases 1 to 8308)
  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: "Sci Rep."; 
            date: "2019-11-20"; volume: "9(1):17188. doi"; pages: " 
            10.1038/s41598-019-53659-4"
COMMENT     Sequence Label: pSLQ1371-sgITGB4-3
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..474
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        476..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             692..872
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    919..1044
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1541..1774
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1959..2003
                     /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    2301..2418
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2671..2704)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2708..2728
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        3022..3325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3326..3529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3563..4159
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3563..3582)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4059..4079
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4169..4222
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      4229..4238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4235..4945
                     /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(4380..4398)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4648..4667
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4965..4998)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5054..5642
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5107..5127)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5525..5536)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5645..5661)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5646..5662)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6171..6351
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6413..7001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6493..6512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7172..7963,7964..8032))
                     /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     7795..7814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8033..8137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8212..8231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
       61 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      121 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      181 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      241 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      301 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      361 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      421 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      481 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      541 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      601 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      661 gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc
      721 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      781 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      841 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
      901 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
      961 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     1021 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     1081 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     1141 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     1201 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     1261 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     1321 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     1381 gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1441 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1501 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1561 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1621 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1681 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1741 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1801 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1861 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1921 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1981 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     2041 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2101 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2161 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2221 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     2281 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2341 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2401 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttagta
     2461 ccgggcccgc tctagagatc cgacgcgcca tctctaggcc cgcgccggcc ccctcgcacg
     2521 gacttgtggg agaagctcgg ctactcccct gccccggtta atttgcatat aatatttcct
     2581 agtaactata gaggcttaat gtgcgataaa agacagataa tctgttcttt ttaatactag
     2641 ctacatttta catgataggc ttggatttct ataacttcgt atagcataca ttatacgaag
     2701 ttataaacag cacaaaagga aactcaccct aactgtaaag taattgtgtg ttttgagact
     2761 ataagtatcc cttggagaac caccttgttg gaggagcgta ggtcctcgca ggtttaagag
     2821 ctatgctgga aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag
     2881 tggcaccgag tcggtgcttt ttttctcgag tactaggatc cattaggcgg ccgcgtggat
     2941 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     3001 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3061 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     3241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     3301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     3361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     3481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     3541 ccgtcagatc cgctagccca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3601 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3661 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3721 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3781 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3841 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3901 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3961 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4021 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4081 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4141 gacccgcaag cccggtgccg gatcgggaga gggcagagga agtctgctaa catgcggtga
     4201 cgtcgaggag aatcctggcc caccggtcgc caccatggtg agcaagggcg aggaggataa
     4261 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     4321 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     4381 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4441 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     4501 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4561 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4621 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4681 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     4741 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     4801 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     4861 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4921 cggcatggac gagctgtaca agtaagaatt cgtcgaggga cctaataact tcgtatagca
     4981 tacattatac gaagttatac atgtttaagg gttccggttc cactaggtac aattcgatat
     5041 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5101 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5161 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5221 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5281 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5341 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5401 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5461 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5521 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5581 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5641 gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca
     5701 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5761 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5821 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     5881 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
     5941 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
     6001 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
     6061 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
     6121 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
     6181 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6241 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6301 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag
     6361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8161 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     8221 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8281 gtagtgcgcg agcaaaattt aagctaca
//
