LOCUS       sequence-318870-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SHIP1 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  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 7080)
  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 Puro shRNA SHIP1 KD1
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    122..355
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             540..584
                     /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        882..1130
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     882..902
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1053..1072
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1223..1340
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1389..1899
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1818..1836
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1921..2520
                     /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(1921..1940)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2417..2437
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2648..2881
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2953..3074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2990..3009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3044..3063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3114..3249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3269..3288)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3270..3288)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3298..3315)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3298..3314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3307..3329)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3456..3911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3753..3774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3937..4041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4042..4110,4111..4902)
                     /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(4260..4279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5073..5661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5562..5581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5815..5832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5949..5970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5985..5990,5991..6008,6009..6015)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6023..6039
                     /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     6028..6050
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6047..6063
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6047..6063
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6082..6102
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6084..6102
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6130..6356
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6357..6537
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6584..6709
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
       61 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
      121 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
      181 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
      241 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
      301 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
      361 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
      421 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
      481 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
      541 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
      601 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
      661 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      721 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
      781 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
      841 atcgatcacg agactagcct cgagcggccg cccccttcac cgagggccta tttcccatga
      901 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
      961 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     1021 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     1081 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggggatctct
     1141 atggcgaagt cctcgaggac ttcgccatag agatcctttt tgaattctcg acctcgagac
     1201 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     1261 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     1321 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggccgc
     1381 ggctcgaggg ggttggggtt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc
     1441 ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc
     1501 ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg
     1561 acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg
     1621 tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca
     1681 atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc
     1741 gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc
     1801 ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc
     1861 ggctccctcg ttgaccgaat caccgacctc tctccccagg gggatccacc ggagcttacc
     1921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     1981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     2041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     2101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     2161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     2221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     2281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     2341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     2401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     2461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     2521 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta
     2581 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     2641 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     2701 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     2761 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     2821 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     2881 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     2941 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3001 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3061 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     3121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3181 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3241 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     3301 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3361 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3421 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3601 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3661 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3781 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3841 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3901 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3961 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4021 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4081 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4141 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4201 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4261 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4321 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4381 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4441 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4501 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4561 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4621 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4681 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4741 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4801 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4861 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4921 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4981 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5041 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5101 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5161 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5221 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5281 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5341 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5401 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5461 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5521 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5581 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5641 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5701 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5761 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5821 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5881 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5941 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6001 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6061 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     6121 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     6181 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     6241 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     6301 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     6361 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6421 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6481 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     6541 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     6601 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     6661 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     6721 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     6781 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     6841 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     6901 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     6961 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     7021 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
//
