LOCUS       sequence-380239-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive lentiviral expression of SLC2A1 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Kim M, Ly SH, Xie Y, Duronio GN, Ford-Roshon D, Hwang JH, Sulahian 
            R, Rennhack JP, So J, Gjoerup O, Talamas JA, Grandclaudon M, Long 
            HW, Doench JG, Sethi NS, Giannakis M, Hahn WC
  TITLE     YAP1 and PRDM14 converge to promote cell survival and tumorigenesis.
  JOURNAL   Dev Cell. 2022 Jan 24;57(2):212-227.e8. doi: 
            10.1016/j.devcel.2021.12.006. Epub 2022 Jan 5.
  PUBMED    34990589
REFERENCE   2  (bases 1 to 7084)
  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.devcel.2021.12.006"; journalName: "Dev Cell"; date: 
            "2022-01-24- 24"; volume: "57"; issue: "2"; pages: "212-227.e8"
COMMENT     Sequence Label: pLKO.1 puro shSLC2A1-1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     464..484
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             695..928
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1000..1121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1037..1056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1091..1110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1161..1296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1316..1335)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1317..1335)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1345..1362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1345..1361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1354..1376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1503..1958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1590..1609)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1800..1821
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1984..2088
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2089..2157,2158..2949)
                     /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     complement(2307..2326)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3120..3708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3609..3628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3862..3879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3996..4017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4032..4037,4038..4055,4056..4062)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4070..4086
                     /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)."
     primer_bind     4075..4097
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4094..4110
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4094..4110
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4129..4149
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4131..4149
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4177..4403
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4404..4584
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4631..4756
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5249..5482
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5667..5711
                     /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"
     promoter        6009..6257
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6009..6029
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6180..6199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6354..6471
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6520..7030
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6949..6967
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7052..7084,1..567)
                     /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(7052..7071)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
       61 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
      121 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
      181 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
      241 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
      301 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
      361 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
      421 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
      481 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
      541 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
      601 aggagcgcac gaccccatgc atcggtacct ttaagaccaa tgacttacaa ggcagctgta
      661 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
      721 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
      781 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      841 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      901 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
      961 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     1021 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1081 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     1141 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1201 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     1261 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     1321 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1381 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1441 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1501 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1561 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1621 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1681 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1741 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1801 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1861 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1921 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     1981 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2041 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2101 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2161 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2221 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2281 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2341 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2401 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2461 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2521 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2581 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2641 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2701 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2761 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2821 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2881 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2941 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3001 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3061 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3781 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3841 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3901 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3961 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4021 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4081 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     4141 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
     4201 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     4261 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     4321 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     4381 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     4441 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     4501 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     4561 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     4621 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     4681 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     4741 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     4801 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     4861 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     4921 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     4981 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     5041 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     5101 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     5161 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     5221 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     5281 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     5341 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     5401 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     5461 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     5521 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     5581 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     5641 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     5701 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     5761 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     5821 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     5881 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     5941 ttcgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
     6001 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     6061 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     6121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     6181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     6241 ggaaaggacg aaacaccggg ccacactatt accatgagaa ctcgagttct catggtaata
     6301 gtgtggcttt ttgaattctc gacctcgaga caaatggcag tattcatcca caattttaaa
     6361 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     6421 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     6481 tacagggaca gcagagatcc actttggccg cggctcgagg gggttggggt tgcgcctttt
     6541 ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg
     6601 cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat
     6661 cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc taagtcggga
     6721 aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac gtctcactag
     6781 taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc gatgggctgt
     6841 ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg
     6901 ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt
     6961 ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa tcaccgacct
     7021 ctctccccag ggggatccac cggagcttac catgaccgag tacaagccca cggtgcgcct
     7081 cgcc
//
