LOCUS       Exported                7435 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  3rd generation lentiviral transfer plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pLV-eGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  AUTHORS   Enomoto M, Bunge MB, Tsoulfas P
  TITLE     A multifunctional neurotrophin with reduced affinity to p75NTR 
            enhances transplanted Schwann cell survival and axon growth after 
            spinal cord injury.
  JOURNAL   Exp Neurol. 2013 Oct;248:170-82. doi: 
            10.1016/j.expneurol.2013.06.013. Epub 2013 Jun 20.
  PUBMED    23792206
REFERENCE   2  (bases 1 to 7435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7435
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             126..986
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2166..2186
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2214..2440
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2441..2621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2668..2793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3286..3519
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3704..3748
                     /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    4015..4131
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        4154..4457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4458..4661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4612..4632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4658..4682
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4751..4760
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4757..5476
                     /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             4757..5473
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4802..4823)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5063..5082)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5410..5431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5492..6080
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5545..5565)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5963..5974)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6167..6400
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6472..6593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6509..6528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6563..6582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6633..6768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6695..6714
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(6788..6807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6789..6807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6817..6834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6817..6833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6826..6848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6975..7430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7062..7081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7272..7293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tccactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     2221 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     2281 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     2341 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     2401 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
     2461 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     2521 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     2581 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     2641 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     2701 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     2761 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     2821 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     2881 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     2941 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     3001 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     3061 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     3121 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     3181 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     3241 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     3301 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     3361 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     3421 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     3481 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     3541 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     3601 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     3661 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     3721 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     3781 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     3841 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     3901 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     3961 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa
     4021 aagaaaaggg gggattgggg ggtatagtgc aggggaaaga atagtagaca taatagcaac
     4081 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag
     4141 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4201 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4261 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4321 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4381 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4441 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4501 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4561 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     4621 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     4681 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagagga
     4741 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4801 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4861 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4921 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4981 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     5041 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     5101 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     5161 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     5221 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     5281 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     5341 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     5401 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     5461 cgagctgtac aagtaaagcg gccgcgtcga caatcaacct ctggattaca aaatttgtga
     5521 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5581 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5641 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5701 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5761 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5821 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5881 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5941 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6001 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6061 cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac
     6121 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     6181 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     6241 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6301 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     6361 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     6421 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     6481 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6541 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6601 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6721 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     6781 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     6841 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6901 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6961 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7021 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7081 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7141 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7201 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7261 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtg
//
