LOCUS       sequence-218950-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miR-9 regulated GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  AUTHORS   Akerblom M, Sachdeva R, Quintino L, Wettergren EE, Chapman KZ, 
            Manfre G, Lindvall O, Lundberg C, Jakobsson J
  TITLE     Visualization and genetic modification of resident brain microglia 
            using lentiviral vectors regulated by microRNA-9.
  JOURNAL   Nat Commun. 2013;4:1770. doi: 10.1038/ncomms2801.
  PUBMED    23612311
REFERENCE   2  (bases 1 to 7897)
  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: "2013"; volume: "4:1770. doi"; pages: " 
            10.1038/ncomms2801"
COMMENT     Sequence Label: pLV.PGK.GFP.miR9T
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(132..200,201..992)
                     /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(350..369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1652..1671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1905..1922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2075..2080,2081..2098,2099..2105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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     2118..2140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2137..2153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2172..2192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2174..2192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2293..2672
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2673..2876
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2827..2847
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2891..3071
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3115..3240
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3733..3966
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4151..4195
                     /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"
     misc_feature    4462..4579
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4626..5136
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5055..5073
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      5152..5161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5158..5160,5161..5163,5164..5877)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5158..5160,5161..5163,5164..5874)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5203..5224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5464..5483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5811..5832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5946..5966)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6364..6375)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6638..6871
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6943..7064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6980..6999)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7034..7053
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7083..7218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7250..7269)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7251..7269)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7279..7296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7279..7295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7288..7310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7440..7895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7527..7546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7737..7758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttggc
     2221 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat
     2281 taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     2341 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     2401 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     2461 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     2521 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     2581 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     2641 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     2701 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2761 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2821 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     2881 tagtgaaccg gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2941 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     3001 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     3061 aatctctagc agtggcgccc gaacagggac ctgaaagcga aagggaaacc agagctctct
     3121 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     3181 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     3241 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     3301 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     3361 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     3421 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     3481 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     3541 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     3601 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     3661 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     3721 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     3781 cgcagcctca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     3841 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     3901 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     3961 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     4021 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     4081 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     4141 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     4201 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     4261 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     4321 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     4381 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     4441 atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg
     4501 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     4561 tacaaaaatt caaaatttta tcgatcacga gactagcctc gagaagcttg atatcgaatt
     4621 cccacggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc
     4681 tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc
     4741 acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg
     4801 cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga
     4861 caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg
     4921 gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcggg gcgcgccgag
     4981 agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg
     5041 ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc
     5101 tccctcgttg accgaatcac cgacctctct ccccaggggg atccaccggt cgccaccatg
     5161 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5221 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5281 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5341 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     5401 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     5461 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     5521 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     5581 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     5641 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     5701 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     5761 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     5821 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     5881 ggccgcgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5941 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6001 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6061 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6121 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6181 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6241 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6301 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6361 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctagagtca
     6421 tacagctaga taaccaaaga cgtatcatac agctagataa ccaaagagct agagtcatac
     6481 agctagataa ccaaagacgt atcatacagc tagataacca aagagctaga gcctcttccg
     6541 cgtcttcgcc ttcccgggtc gagctcggta cctttaagac caatgactta caaggcagct
     6601 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6661 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6721 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6781 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6841 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6901 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6961 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7021 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     7081 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     7141 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     7201 gaggcttttt tggaggccta ggcttttgcg tcgagacgta cccaattcgc cctatagtga
     7261 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     7321 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     7381 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga
     7441 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7501 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7561 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7621 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7681 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7741 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     7801 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7861 cgcgaatttt aacaaaatat taacgtttac aatttcc
//
