LOCUS       sequence-180322-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector with constitutive expression of shRNA targeting 
            human FOXM1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  TITLE     FOXM1 toolbox
  JOURNAL   Unpublished
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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLKO-FOXM1-shRNA-81
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             130..729
                     /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(130..149)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     626..646
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             857..1090
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1162..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1199..1218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1253..1272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1323..1458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1478..1497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1479..1497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1507..1524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1507..1523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1516..1538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1665..2120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1752..1771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1962..1983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2146..2250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2251..2319,2320..3111)
                     /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(2469..2488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3282..3870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3771..3790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4024..4041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4158..4179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4194..4199,4200..4217,4218..4224)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4232..4248
                     /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     4237..4259
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4256..4272
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4256..4272
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4291..4311
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4293..4311
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4339..4565
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4566..4746
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4793..4918
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5411..5644
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5829..5873
                     /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        6171..6419
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6171..6191
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6342..6361
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6516..6633
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(6682..7084,1..108)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg
       61 tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccaccg
      121 gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
      181 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
      241 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
      301 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
      361 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
      421 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
      481 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
      541 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
      601 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
      661 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
      721 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
      781 gcatcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
      841 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
      901 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      961 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     1021 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     1081 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     1141 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     1201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1261 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     1321 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1381 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1441 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     1501 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1561 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1621 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     1681 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1741 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1801 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1861 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1921 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1981 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2041 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2101 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4081 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4141 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     4201 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4261 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     4321 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     4381 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     4441 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     4501 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     4561 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     4621 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     4681 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     4741 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     4801 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     4861 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     4921 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     4981 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     5041 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     5101 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     5161 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     5221 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     5281 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     5341 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     5401 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     5461 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     5521 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     5581 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     5641 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     5701 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     5761 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     5821 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     5881 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     5941 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     6001 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     6061 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     6121 ctcgacggta tcgatcacga gactagcctc gagcggccgc ccccttcacc gagggcctat
     6181 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     6241 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     6301 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     6361 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     6421 gaggaccact ttccctactt tactcgagta aagtagggaa agtggtcctt ttttgaattc
     6481 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     6541 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     6601 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     6661 ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt
     6721 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
     6781 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
     6841 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
     6901 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
     6961 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
     7021 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     7081 ggta
//
