LOCUS       sequence-235393-        8308 bp    DNA     circular SYN 15-APR-2025
DEFINITION  sgITGB1-3 sequence: GTTCAGTGAATGGGAACAACG. 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 Apr 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-sgITGB1-3
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        90..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             631..1227
                     /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(631..650)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1127..1147
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1237..1290
                     /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      1297..1306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1303..2013
                     /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(1448..1466)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1716..1735
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2033..2066)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2122..2710
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2175..2195)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2593..2604)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2713..2729)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2714..2730)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3239..3419
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3481..4069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3561..3580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4240..5031,5032..5100))
                     /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     4863..4882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5101..5205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5280..5299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5471..5850
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5852..6050
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6068..6248
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6295..6420
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6917..7150
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7335..7379
                     /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    7677..7794
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(8047..8080)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     8084..8104
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 gcgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      601 ttagtgaacc gtcagatccg ctagcccacc atgaccgagt acaagcccac ggtgcgcctc
      661 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
      721 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
      781 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
      841 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
      901 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
      961 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     1021 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     1081 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     1141 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     1201 tggtgcatga cccgcaagcc cggtgccgga tcgggagagg gcagaggaag tctgctaaca
     1261 tgcggtgacg tcgaggagaa tcctggccca ccggtcgcca ccatggtgag caagggcgag
     1321 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     1381 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     1441 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     1501 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     1561 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1621 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1681 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1741 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     1801 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     1861 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     1921 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     1981 tccaccggcg gcatggacga gctgtacaag taagaattcg tcgagggacc taataacttc
     2041 gtatagcata cattatacga agttatacat gtttaagggt tccggttcca ctaggtacaa
     2101 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2161 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2221 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2281 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2341 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2401 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2461 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2521 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2581 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2641 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2701 gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac
     2761 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     2821 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     2881 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     2941 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     3001 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
     3061 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt
     3121 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
     3181 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
     3241 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     3301 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3361 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     3421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5221 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     5281 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     5341 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     5401 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     5461 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     5521 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     5581 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5641 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     5701 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     5761 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     5821 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     5881 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     5941 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     6001 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
     6061 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6121 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6181 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6241 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     6301 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     6361 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     6421 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     6481 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     6541 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     6601 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     6661 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     6721 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     6781 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     6841 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     6901 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     6961 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     7021 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     7081 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     7141 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     7201 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     7261 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     7321 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     7381 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     7441 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     7501 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     7561 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     7621 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     7681 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     7741 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     7801 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gagatccgac
     7861 gcgccatctc taggcccgcg ccggccccct cgcacggact tgtgggagaa gctcggctac
     7921 tcccctgccc cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc
     7981 gataaaagac agataatctg ttctttttaa tactagctac attttacatg ataggcttgg
     8041 atttctataa cttcgtatag catacattat acgaagttat aaacagcaca aaaggaaact
     8101 caccctaact gtaaagtaat tgtgtgtttt gagactataa gtatcccttg gagaaccacc
     8161 ttgttggttc agtgaatggg aacaacggtt taagagctat gctggaaaca gcatagcaag
     8221 tttaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     8281 ctcgagtact aggatccatt aggcggcc
//
