LOCUS       sequence-139182-        6767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral shRNA vector. Derivative of pMKO.1 puro (Addgene plasmid 
            #8452) with GFP instead of puromycin resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6767)
  TITLE     Viral vectors.
REFERENCE   2  (bases 1 to 6767)
  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"
COMMENT     Sequence Label: pMKO.1 GFP
FEATURES             Location/Qualifiers
     source          1..6767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(197..216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(238..567)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      252..387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     708..730
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     LTR             833..1258
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     CDS             complement(1410..2129)
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     2134..2150
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2135..2151
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2160..2489)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2174..2309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2550..2798)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2608..2627)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2778..2798)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(2834..3250)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2850..2866)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2882..2904)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3451..3650
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3713..3888
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     promoter        complement(3889..4092)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3918..3938)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4093..4472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4572..4590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4868..4972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4973..5041,5042..5833)
                     /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(5191..5210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6004..6592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6493..6512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
       61 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      121 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      181 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag taagcttttt
      241 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
      301 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
      361 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
      421 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      481 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      541 ggactttcca caccctaact gacacacatt ccacagccgg atctacactc cgctatcgct
      601 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      661 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      721 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagccagctt acctcccggt
      781 ggtgggtcgg tggtccctgg gcaggggtct cccgatcccg gacgagcccc caaatgaaag
      841 acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac agcgagacca
      901 caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg cgactcagtc
      961 aatcggagga ctggcgcccc gagtgagggg ttgtgggctc ttttattgag ctcggggagc
     1021 agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata aggcacaggg
     1081 tcatttcagg tccttggggc accctggaaa catctgatgg ttctctaggg tcgaccctgt
     1141 tccatctgtt cctgaccttg atctgaactt ctctattctc agttatgtat ttttccatgc
     1201 cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt ctttcattcc
     1261 cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt actctatagg
     1321 cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata tcctgtcttt
     1381 aacaaattgg actaatcgaa ggtaccctat tacacccact cgtgcaggct gcccaggggc
     1441 ttgcccaggc tggtcagctg ggcgatggcg gtctcgtgct gctccacgaa gccgccgtcc
     1501 tccacgtagg tcttctccag gcggtgctgg atgaagtggt actcggggaa gtccttcacc
     1561 acgcccttgc tcttcatcag ggtgcgcatg tggcagctgt agaacttgcc gctgttcagg
     1621 cggtacacca ggatcacctg gcccaccagc acgccgtcgt tcatgtacac cacctcgaag
     1681 ctgggctgca ggccggtgat ggtcttcttc atcacggggc cgtcgttggg gaagttgcgg
     1741 cccttgtact ccacgcggta cacgaacatc tcctcgatca ggttgatgtc gctgcggatc
     1801 tccaccaggc cgccgtcctc gtagcgcagg gtgcgctcgt acacgaagcc ggcggggaag
     1861 ctctggatga agaagtcgct gatgtcctcg gggtacttgg tgaaggtgcg gttgccgtac
     1921 tggaaggcgg ggctcaggat gtcgaaggcg aagggcaggg gggcgccctt ggtcacgcgg
     1981 atctgcacca gctggttgcc gaacaggatg ttgcccttgc cgcagccctc catggtgaac
     2041 acgtggttgt tcaccacgcc ctccaggttc accttgaagc tcatgatctc ctgcaggccg
     2101 gtgttcttca ggatctgctt gctcaccatg gtctcgaggt cgacggtatc gataagcttt
     2161 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     2221 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2281 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     2341 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     2401 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     2461 ggggactttc cacaccctaa ctgacacaca ttccacaggg tcgaccactg tgctggcgaa
     2521 ttccatatgg gagtactagt acgcgtaccg gtgtttcgtc ctttccacaa gatatataaa
     2581 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
     2641 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
     2701 aatatctttg tgtttgcagt caaattaatt ctaattatct ctctaacagc cttgtatcgt
     2761 atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac cttgtcgaca
     2821 gatcttcaat tccggcgcct agagaaggag tgagggctgg ataaagggag gatcgaggcg
     2881 gggtcgaacg aggaggttca agggggagag acggggcgga tggaggaaga ggaggcggag
     2941 gcttagggtg tacaaagggc ttgacccagg gaggggggtc aaaagccaag gcttcccagg
     3001 tcacgatgta ggggacctgg tctgggtgtc catgcgggcc aggtgaaaag accttgatct
     3061 taacctgggt gatgaggtct cggttaaagg tgccgtctcg cggccatccg acgttaaagg
     3121 ttggccattc tgcagagcag aaggtaaccc aacgtctctt cttgacatct accgactggt
     3181 tgtgagcgat ccgctcgaca tctttccagt gacctaaggt caaacttaag ggagtggtaa
     3241 cagtctggcc ctaattttca gacaaataca gaaacacagt cagacagaga caacacagaa
     3301 cgatgctgca gcgctgcagc agacaagacg cgcggcttcg gttccaaacc gaaagcaaaa
     3361 attcagacgg aggcgggaac tgttttaggt tctcgtctcc taccagaacc acatatcctg
     3421 acggggtcgg attccacatc gactcccttc ctcaggtcgg gccacaaaaa cggcccccaa
     3481 agtccctggg acgtctccca gggttgcggc cgggtgttca gaactcgtca gttccaccac
     3541 gggtccgcca gatacagagc tagttagcta actagtacag acgcaggcgc ataacatcaa
     3601 acatagacac tagacaatcg gacagacaca gataagttgc tggccagctt acctcccggt
     3661 ggtgggtcgg tggtccctgg gcaggggtct ccaaatcccg gacgagcccc caaatgaaag
     3721 acccccgtcg tgggtagtca atcactcaga ggagaccctc ccaaggaaca gcgagaccac
     3781 gattcggatg caaacagcaa gaggctttat tgggaatacg ggtacccggg cgacgcagtc
     3841 tatcggaaga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag ctctgcttat
     3901 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     3961 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     4021 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     4081 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     4141 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     4201 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     4261 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     4321 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     4381 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     4441 ttactattaa taactagtca ataatcaatg tcaacatggc ggtaatgttg gacatgagcc
     4501 aatataaatg tacatattat gatatggata caacgtatgc aatggccaag ctaattcttg
     4561 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4621 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4741 ataattaagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4801 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     4861 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4921 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4981 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5041 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5101 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5161 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5221 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5281 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5341 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5401 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5461 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5521 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5581 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5641 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5701 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5761 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5821 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5881 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5941 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6001 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6061 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6721 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaag
//
