LOCUS       Exported                6114 bp ds-DNA     linear   SYN 24-MAY-2021
DEFINITION  to package wild type EnvA psudotyped lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    pCAGGS/ES-EnvA-VSVg-WPRE-pA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6114)
  AUTHORS   Sakurai K, Zhao S, Takatoh J, Rodriguez E, Lu J, Leavitt AD, Fu M, 
            Han BX, Wang F
  TITLE     Capturing and Manipulating Activated Neuronal Ensembles with CANE 
            Delineates a Hypothalamic Social-Fear Circuit.
  JOURNAL   Neuron. 2016 Nov 23;92(4):739-753. doi: 
            10.1016/j.neuron.2016.10.015. Epub 2016 Oct 27.
  PUBMED    27974160
REFERENCE   2  (bases 1 to 6114)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 24, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6114
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1971..2559
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2024..2044)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2442..2453)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2591..3067
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3195..3214)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3260..3315
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3314..3333)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3676..3692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3676..3692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3689..3711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3700..3716
                     /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)."
     promoter        complement(3724..3754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3769..3790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3849..4044
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3895..4030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3957..3976
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4050..4184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4087..4106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4141..4160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4252..4269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4423..5011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4503..4522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5182..6042)
                     /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     5805..5824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6043..6114)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacataccct accgagaact cagagagtcg ttggaagacg ggaaggaagc ccgacgactg
       61 agcggtccac cccagacgtg gttctggtcg cccggaggat caagtatgga agccgtcata
      121 aaggcatttc tgactgggca ccctggaaag gtgagcaaga aggactccaa gaagaagccg
      181 ccagcaacaa gcaagaaaga cccggagaag acgcccttgc tgccaacgag agttaattat
      241 attattggtg tcctggtctt gtgtgaggta acgggggtaa gagctgatgt ccacttactc
      301 gagcagccag ggaacctttg gattacatgg gccaaccgta caggccaaac ggatttttgc
      361 ctctctacac agtcagccac ctcccctttt caaacatgtt tgataggtat cccgtcccct
      421 atttccgagg gtgattttaa gggatatgtt tctgatacaa attgcaccac cttgggaact
      481 gatcggttag tctcgtcagc cgactttact ggcggacctg acaacagtac caccctcact
      541 tatcggaagg tctcatgctt gttgttaaag ctgaatgtct ctatgtggga tgagccacct
      601 gaactacagc tgttaggttc ccagtctctc cctaacatta ctaatattgc tcagatttcc
      661 ggtataaccg ggggatgcgt aggcttcaga ccacaagggg ttccttggta tctaggttgg
      721 tctagacagg aggccacgcg gtttctcctt agacacccct ctttctctaa atccacggaa
      781 ccgtttacgg tggtgacagc ggataggcac aatcttttta tggggagtga gtactgcggt
      841 gcatatggct acagattttg gaacatgtat aactgctcac aggtggggcg gcagtaccgc
      901 tgtggtaatg cgcgcagccc ccgcccgggt cttcctgaaa tccagtgtac aaggagagga
      961 ggcaaatggg ttaatcaatc acaggaaatt aatgagtcgg agccgttcag ctttacggtg
     1021 aactgtacag ctagtagttt gggtaatgcc agtgggtgtt gcggaaaagc aggcacgatt
     1081 ctcccgggaa agtgggtcga cagcacacaa ggtagtttca ccaaaccaaa agcgctacca
     1141 cccgcaattt tcctcatttg tggggatcgc gcatggcaag gaattcccag tcgtccggta
     1201 gggggcccct gctatttagg caagcttacc atgttagcac ctaagcatac agatattctc
     1261 aaggtgcttg tcaattcatc gcggacaggt ataagacgta aacgaagcac ctcacacctg
     1321 gatgatacat gctcagatga agtgcagctt tggggtccta cagcaagaat ctttgcatct
     1381 atcctagccc cgggggtagc agctgcgcaa gccttaagag aaattgagag actagcctgt
     1441 tggtccgtta aacaggctaa cttgacaaca tcactcctcg gggacttatt ggatgatgtc
     1501 acgagtattc gacacgcggt cctgcagaac cgagcggcta ttgacttctt gctcctagct
     1561 cacggccatg gctgtgagga cgttgccgga atgtgctgtt tcaatttgag tgatcacagt
     1621 gagtctatac agaagaagtt ccagctaatg aaggaacatg tcaataagat cggcgtggac
     1681 agcgacccaa ttggaagttg gctgcgagga ctattcgggg gaataggaga atgggccgtt
     1741 catttgctga aaggactgct tttggggctt gtagttattt tgttgctagt agtgtgcctg
     1801 ccttgccttt tgctcgagcg agttggtatt tatctttgca ttaaattaaa gcacaccaag
     1861 aaaagacaga tttatacaga catagagatg aaccgacttg gaaggtaacg gatccttcga
     1921 aaccggtgct agcgcgcccc tgcagggaat tcgatatcaa gcttatcgat aatcaacctc
     1981 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2041 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2101 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2161 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2221 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2281 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2341 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     2401 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2461 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2521 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     2581 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2641 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2701 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2761 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     2821 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     2881 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     2941 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3001 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3061 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcgagc tcaattcact
     3121 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     3181 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3241 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3301 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3361 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3421 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3481 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3541 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     3601 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     3661 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3841 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     4081 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4141 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     4201 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4261 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4321 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4381 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4441 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4501 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4561 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4621 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4681 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4741 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4801 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4861 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4921 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4981 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5041 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5101 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5161 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5221 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5281 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5341 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5401 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5461 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5521 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5581 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5641 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5701 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5761 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5821 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5881 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5941 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6001 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6061 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatg
//
