LOCUS       sequence-361631-        7767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with BaeI site to allow cloning of custom sgRNA into vector 
            containing Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7767)
  AUTHORS   Cai H, Chew SK, Li C, Tsai MK, Andrejka L, Murray CW, Hughes NW, 
            Shuldiner EG, Ashkin EL, Tang R, Hung KL, Chen LC, Lee SYC, Yousefi 
            M, Lin WY, Kunder CA, Cong L, McFarland CD, Petrov DA, Swanton C, 
            Winslow MM
  TITLE     A functional taxonomy of tumor suppression in oncogenic KRAS-driven 
            lung cancer.
  JOURNAL   Cancer Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325.
  PUBMED    33608386
REFERENCE   2  (bases 1 to 7767)
  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: "Cancer 
            Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325."
COMMENT     Sequence Label: Lenti-BaeI-Pgk-Cre
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        310..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        691..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     844..864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             907..1087
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1134..1259
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1756..1989
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2174..2218
                     /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    2516..2633
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2715..2963
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2715..2735
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2886..2905
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2992..3067
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3120..3619
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3155..3178)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3552..3571
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3623..3643
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3644..4672
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    4728..5316
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4781..4801)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5199..5210)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5319..5335)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5320..5336)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5845..6025
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6087..6675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6167..6186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6846..7637,7638..7706))
                     /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     7469..7488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7707..7767,1..44))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
       61 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
      121 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
      181 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
      241 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      301 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      361 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
      901 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      961 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1021 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1081 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1141 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1201 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1261 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1321 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1381 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1441 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1501 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1561 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
     1621 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1681 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1741 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1801 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1861 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1921 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1981 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     2041 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     2101 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     2161 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     2221 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     2281 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     2341 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     2401 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     2461 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     2521 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     2581 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     2641 attacaggga cagcagagat ccagtttggt tagtaccggg cccgctctag ccctgcagga
     2701 actgtttaaa ctgtgagggc ctatttccca tgattccttc atatttgcat atacgataca
     2761 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2821 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2881 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2941 tcttgtggaa aggaccaaac accgcgaggt attaccggcg tatcatccgc ggttttagag
     3001 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     3061 tcggtgcttt tttgtttggc gcgcctatgg cttaattaac atggcggccg cattctaccg
     3121 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     3181 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     3241 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     3301 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     3361 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     3421 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     3481 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     3541 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     3601 ctcatctccg ggacccggga tgcccaagaa gaagaggaag gtgtccaatt tactgaccgt
     3661 acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct
     3721 gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc
     3781 cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga
     3841 acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac
     3901 tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc
     3961 aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc
     4021 cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc
     4081 actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc
     4141 ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg
     4201 tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc
     4261 aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt
     4321 ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt
     4381 tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc
     4441 aactcatcga ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc
     4501 tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc
     4561 ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa
     4621 cctggatagt gaaacagggg caatggtgcg cctgctggaa gatggcgatt agccatgttt
     4681 aagggttccg gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg
     4741 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4801 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4861 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4921 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4981 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5041 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5101 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5161 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5221 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5281 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga
     5341 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     5401 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     5461 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     5521 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     5581 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
     5641 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
     5701 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
     5761 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
     5821 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
     5881 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5941 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6001 tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac
     6061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7741 cagggttatt gtctcatgag cggatac
//
