LOCUS       sequence-198940-        7084 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Suppression of ABCB7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Alvarez SW, Sviderskiy VO, Terzi EM, Papagiannakopoulos T, Moreira 
            AL, Adams S, Sabatini DM, Birsoy K, Possemato R
  TITLE     NFS1 undergoes positive selection in lung tumours and protects cells
            from ferroptosis.
  JOURNAL   Nature. 2017 Nov 22. doi: 10.1038/nature24637.
  PUBMED    29168506
REFERENCE   2  (bases 1 to 7084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Nov 22. doi: 10.1038/nature24637."
COMMENT     Sequence Label: pLKO.1P shABCB7_2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..523
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     442..460
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             545..1144
                     /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(545..564)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1041..1061
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1272..1505
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1577..1698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1614..1633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1668..1687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1738..1873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1893..1912)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1894..1912)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1922..1939)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1922..1938)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1931..1953)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2080..2535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2167..2186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2377..2398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2561..2665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2666..2734,2735..3526)
                     /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(2884..2903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3697..4285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4186..4205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4439..4456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4573..4594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4609..4614,4615..4632,4633..4639)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4647..4663
                     /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     4652..4674
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4671..4687
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4671..4687
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4706..4726
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4708..4726
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4754..4980
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4981..5161
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5208..5333
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5826..6059
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6244..6288
                     /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        6586..6834
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6586..6606
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6757..6776
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6931..7048
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg
       61 acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac
      121 attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc
      181 ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg
      241 gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg
      301 agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg
      361 cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg
      421 ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc
      481 agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggagct
      541 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
      601 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
      661 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      721 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      781 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      841 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      901 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      961 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     1021 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     1081 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     1141 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     1201 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     1261 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     1321 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     1381 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     1441 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     1501 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     1561 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1621 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1681 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     1741 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1801 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     1861 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     1921 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     1981 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2041 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     2101 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2161 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2221 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2281 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2341 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2401 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2461 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2521 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2641 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2701 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2761 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2821 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2881 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2941 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3001 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3061 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3121 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3181 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3241 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3301 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3361 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3421 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3481 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4381 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4441 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4501 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4561 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4621 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4681 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     4741 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     4801 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     4861 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     4921 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     4981 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5041 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5101 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5161 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     5221 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     5281 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     5341 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     5401 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     5461 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     5521 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     5581 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     5641 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     5701 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     5761 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     5821 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     5881 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     5941 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     6001 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     6061 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     6121 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     6181 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     6241 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     6301 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     6361 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     6421 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     6481 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     6541 cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg cctatttccc
     6601 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     6661 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     6721 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     6781 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggcat
     6841 ttatcttgct ggtcaaactc gagtttgacc agcaagataa atgctttttg aattctcgac
     6901 ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     6961 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     7021 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact
     7081 ttgg
//
