LOCUS       sequence-180323-        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  JOURNAL   Unpublished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            "Unpublished"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-84
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             88..687
                     /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(88..107)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     584..604
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             815..1048
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1120..1241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1157..1176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1211..1230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1281..1416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1436..1455)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1437..1455)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1465..1482)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1465..1481)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1474..1496)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1623..2078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1710..1729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1920..1941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2104..2208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2209..2277,2278..3069)
                     /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(2427..2446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3240..3828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3729..3748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3982..3999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4116..4137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4152..4157,4158..4175,4176..4182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4190..4206
                     /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     4195..4217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4214..4230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4214..4230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4249..4269
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4251..4269
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4297..4523
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4524..4704
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4751..4876
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5369..5602
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5787..5831
                     /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        6129..6377
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6129..6149
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6300..6319
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6474..6591
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(6640..7084,1..66)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     join(7069..7084,1..3)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
       61 ccccaggggg atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc
      121 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
      181 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
      241 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
      301 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
      361 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
      421 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
      481 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
      541 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
      601 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
      661 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
      721 aggagcgcac gaccccatgc atcggtacct ttaagaccaa tgacttacaa ggcagctgta
      781 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
      841 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
      901 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      961 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1021 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     1081 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     1141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     1261 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1321 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     1381 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     1441 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1501 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1561 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1621 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1681 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1801 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1921 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1981 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2041 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     2101 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2161 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2221 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2281 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2341 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2401 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2461 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2521 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2581 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2641 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2701 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2761 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2821 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2881 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2941 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3001 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3061 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3121 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3181 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3241 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3301 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3361 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3421 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3481 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3541 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3601 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3661 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3721 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3781 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3841 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3901 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3961 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4021 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4081 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4141 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4201 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     4261 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
     4321 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     4381 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     4441 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     4501 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     4561 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     4621 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     4681 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     4741 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     4801 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     4861 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     4921 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     4981 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     5041 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     5101 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     5161 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     5221 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     5281 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     5341 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     5401 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     5461 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     5521 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     5581 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     5641 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     5701 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     5761 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     5821 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     5881 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     5941 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     6001 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     6061 ttcgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
     6121 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     6181 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     6241 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     6301 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     6361 ggaaaggacg aaacaccggt tgcagggtgg tccgtgtaaa ctcgagttta cacggaccac
     6421 cctgcaattt ttgaattctc gacctcgaga caaatggcag tattcatcca caattttaaa
     6481 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     6541 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     6601 tacagggaca gcagagatcc actttggccg cggctcgagg gggttggggt tgcgcctttt
     6661 ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg
     6721 cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat
     6781 cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc taagtcggga
     6841 aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac gtctcactag
     6901 taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc gatgggctgt
     6961 ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg
     7021 ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt
     7081 ctgc
//
