LOCUS       sequence-318818-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  INPP4B shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, 
            McGarry L, Shanks E, Mostov KE, Bryant DM
  TITLE     The phospholipid PI(3,4)P2 is an apical identity determinant.
  JOURNAL   Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.
  PUBMED    30487552
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: "Nat 
            Commun."; date: "2018-11-28"; volume: "9(1):5041. doi"; pages: " 
            10.1038/s41467-018-07464-8"
COMMENT     Sequence Label: pLKO.1 Hygro shRNA INPP4B KD2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..513
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     432..450
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             535..1134
                     /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(535..554)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1031..1051
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1262..1495
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1567..1688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1604..1623)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1658..1677
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1728..1863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1883..1902)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1884..1902)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1912..1929)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1912..1928)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1921..1943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2070..2525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2157..2176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2367..2388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2551..2655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2656..2724,2725..3516)
                     /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(2874..2893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3687..4275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4176..4195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4429..4446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4563..4584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4599..4604,4605..4622,4623..4629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4637..4653
                     /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     4642..4664
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4661..4677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4661..4677
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4696..4716
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4698..4716
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4744..4970
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4971..5151
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5198..5323
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5816..6049
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6234..6278
                     /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        6576..6824
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6576..6596
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6747..6766
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6921..7038
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc
       61 tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg
      121 tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt
      181 cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa
      241 acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca
      301 gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc
      361 agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc
      421 ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc
      481 ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggagct taccatgacc
      541 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
      601 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
      661 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
      721 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
      781 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
      841 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
      901 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
      961 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1021 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1081 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     1141 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc ggtaccttta
     1201 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     1261 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     1321 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     1381 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     1441 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     1501 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     1561 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1681 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     1741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1801 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1861 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     1921 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1981 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2041 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     2101 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2161 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2221 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2281 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2341 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2401 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2521 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2761 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2821 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2881 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2941 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3001 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3061 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3121 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3181 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3241 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3301 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3361 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3421 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3481 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4321 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4381 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4441 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4501 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4561 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     4621 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4681 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct
     4741 taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac
     4801 atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg
     4861 atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga accactgaat
     4921 tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct
     4981 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5041 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5101 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     5161 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     5221 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     5281 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     5341 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     5401 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     5461 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     5521 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     5581 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     5641 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     5701 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     5761 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     5821 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     5881 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     5941 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     6001 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     6061 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     6121 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     6181 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     6241 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     6301 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     6361 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     6421 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     6481 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat
     6541 cacgagacta gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt
     6601 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
     6661 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     6721 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     6781 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggctc atgacaccgt
     6841 tcaaaccctc gagggtttga acggtgtcat gagctttttg aattctcgac ctcgagacaa
     6901 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     6961 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     7021 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggccgcgg
     7081 ctcg
//
