LOCUS       Exported                7902 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  miR-9 regulated GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    pLV.PGK.GFP.miR9T
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7902)
  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 7902)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7902
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             583..763
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    810..935
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1428..1661
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1846..1890
                     /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    2157..2274
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2320..2830
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2749..2767
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      2846..2855
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2852..3571
                     /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             2852..3568
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2897..2918)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3158..3177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3505..3526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3640..3660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4058..4069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4332..4565
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4637..4758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4674..4693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4728..4747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4798..4933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4860..4879
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4953..4972)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4954..4972)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4982..4999)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4982..4998)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4991..5013)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5140..5595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5227..5246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5437..5458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5621..5725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5726..6586
                     /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(5944..5963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6757..7345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7246..7265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7499..7516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7633..7654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7669..7699
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7707..7723
                     /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     7712..7734
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7731..7747
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7731..7747
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7766..7786
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7768..7786
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        7887..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctcg tttagtgaac cggggtctct ctggttagac
      601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
      661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
      721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
      781 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
      841 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
      901 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
      961 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1021 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1081 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1141 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1201 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1261 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1321 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1381 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     1441 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     1501 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     1561 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     1621 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     1681 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     1741 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     1801 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     1861 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     1921 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     1981 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2041 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2101 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt
     2161 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2221 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttatcgat
     2281 cacgagacta gcctcgagaa gcttgatatc gaattccacg gggttggggt tgcgcctttt
     2341 ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg
     2401 cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat
     2461 cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc taagtcggga
     2521 aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac gtctcactag
     2581 taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc gatgggctgt
     2641 ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg
     2701 ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt
     2761 ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa tcaccgacct
     2821 ctctccccag ggggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     2881 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2941 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3001 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3061 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3121 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3181 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3241 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3301 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3361 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3421 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3481 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3541 cactctcggc atggacgagc tgtacaagta aagcggccgc gtcgacaatc aacctctgga
     3601 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3661 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3721 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3781 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3841 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3901 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3961 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4021 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4081 tccttcccgc ggcctgctgc cggctctaga gtcatacagc tagataacca aagacgtatc
     4141 atacagctag ataaccaaag agctagagtc atacagctag ataaccaaag acgtatcata
     4201 cagctagata accaaagagc tagagcctct tccgcgtctt cgccttcccg ggtcgagctc
     4261 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     4321 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     4381 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     4441 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     4501 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4561 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     4621 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4741 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     4801 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4861 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4921 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     4981 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5041 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5101 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     5161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5341 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5401 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5461 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5521 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5581 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5701 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5761 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5821 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5881 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5941 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6001 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6061 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6121 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6181 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6241 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6301 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6361 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6421 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6481 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6541 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6601 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6661 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6721 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6781 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6841 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6901 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6961 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7021 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7081 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7141 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7201 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7261 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7321 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7381 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7441 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7501 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7561 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7621 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     7681 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7741 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     7801 gctgcaagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     7861 ctcatgtcca acattaccgc catgttgaca ttgattattg ac
//
