LOCUS       sequence-254625-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shLuc (Target TTACGCTGAGTACTTCGA), silence LUC gene, blasticidin 
            selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  AUTHORS   Sandra Martha Gomes Dias
  TITLE     HuR (ELAVL1) silencing sensitizes triple-negative breast cancer to 
            glutaminase-targeted therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6816)
  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            "Unpublished"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.1-blast shLuc
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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        1606..1854
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1606..1626
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1777..1796
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1950..2067
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2116..2626
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2545..2563
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2638..3036
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3108..3341
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3413..3534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3450..3469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3504..3523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3574..3709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3729..3748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3730..3748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3758..3775)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3758..3774)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3767..3789)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3916..4371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4003..4022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4213..4234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4397..4501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4502..4570,4571..5362)
                     /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(4720..4739)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5533..6121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6022..6041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6275..6292
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6409..6430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6445..6450,6451..6468,6469..6475)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6483..6499
                     /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     6488..6510
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6507..6523
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6507..6523
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6542..6562
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6544..6562
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6590..6816
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg cctatttccc
     1621 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     1681 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     1741 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     1801 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggctta
     1861 cgctgagtac ttcgacctga cccatcgaag tactcagcgt aagtttttga attctcgacc
     1921 tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     1981 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2041 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     2101 tggccgcggc tcgagggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca
     2161 gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg
     2221 cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc
     2281 cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg
     2341 ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca
     2401 gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg
     2461 gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt
     2521 gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc
     2581 ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg atccaccatg
     2641 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     2701 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     2761 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     2821 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     2881 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     2941 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     3001 gaattgctgc cctctggtta tgtgtgggag ggctaaggta cctttaagac caatgactta
     3061 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     3121 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     3181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     3241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     3301 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     3361 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     3421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3541 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3601 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3661 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     3721 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     3781 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3841 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3901 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3961 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4021 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4081 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4141 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4201 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4261 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4321 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     4381 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4441 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4501 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4561 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4621 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4681 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4741 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4801 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4861 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4921 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4981 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5041 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5101 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5161 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5221 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5281 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5341 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5401 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5461 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5521 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5581 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5641 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5701 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5761 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5821 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5881 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5941 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6001 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6061 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6121 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6181 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6241 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6301 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6361 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6421 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6481 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     6541 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat
     6601 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
     6661 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
     6721 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
     6781 gatattgtat ttaagtgcct agctcgatac ataaac
//
