LOCUS       sequence-227288-        6817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shGFP control, silence GFP gene, blasticidin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  TITLE     Dias lab diverse plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6817)
  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            "Unpublished"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 shGFP
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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        1833..2081
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1833..1853
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2004..2023
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2178..2295
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2344..2854
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2773..2791
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2866..3264
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3336..3569
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3641..3762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3678..3697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3732..3751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3802..3937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3957..3976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3958..3976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3986..4003)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3986..4002)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3995..4017)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4144..4599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4231..4250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4441..4462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4625..4729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4730..4798,4799..5590)
                     /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(4948..4967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5761..6349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6250..6269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6503..6520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6637..6658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6673..6678,6679..6696,6697..6703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6711..6727
                     /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     6716..6738
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6735..6751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6735..6751
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6770..6790
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6772..6790
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgatcac
     1801 gagactagcc tcgagcggcc gcccccttca ccgagggcct atttcccatg attccttcat
     1861 atttgcatat acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca
     1921 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     1981 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     2041 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggcaagctg accctgaagt
     2101 tcatctcgag atgaacttca gggtcagctt gtttttgaat tctcgacctc gagacaaatg
     2161 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2221 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2281 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg gccgcggctc
     2341 gagggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg
     2401 ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac
     2461 gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct
     2521 tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca
     2581 aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc
     2641 agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag
     2701 cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt
     2761 cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc
     2821 cctcgttgac cgaatcaccg acctctctcc ccagggggat ccaccatggc caagcctttg
     2881 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     2941 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     3001 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     3061 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     3121 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     3181 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     3241 tctggttatg tgtgggaggg ctaaggtacc tttaagacca atgacttaca aggcagctgt
     3301 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     3361 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     3421 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3481 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3541 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     3601 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     3661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     3781 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3841 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3901 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     3961 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4021 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4081 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4141 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4201 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4261 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4321 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4381 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4441 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4501 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4561 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4621 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4681 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4741 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4801 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4861 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4921 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4981 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5041 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5101 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5161 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5221 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5281 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5341 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5401 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5461 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5521 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5581 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5641 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5881 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6421 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6481 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6541 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6601 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6661 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6721 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     6781 tcactaaagg gaacaaaagc tggagctgca agcttaa
//
