LOCUS       sequence-170230-        7920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to package wild type EnvA psudotyped lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7920)
  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 7920)
  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"; doi: 
            "10.1016/j.neuron.2016.10.015"; journalName: "Neuron"; date: 
            "2016-11-23- 23"; volume: "92"; issue: "4"; pages: "739-753"
COMMENT     Sequence Label: pCAGGS/ES-EnvA-VSVg-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     misc_feature    3714..4302
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3767..3787)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4185..4196)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4334..4810
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4533..4552)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4944..4963)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5009..5064
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5063..5082)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5423..5439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5423..5439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5436..5458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5447..5463
                     /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(join(5471..5477,5478..5495,5496..5501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5516..5537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5596..5791
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5642..5777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5797..5931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5834..5853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5888..5907
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5999..6016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6170..6758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6250..6269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6929..7720,7721..7789))
                     /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     7552..7571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7790..7894)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtacc cgtgcaggga
     1741 gccaacatac cctaccgaga actcagagag tcgttggaag acgggaagga agcccgacga
     1801 ctgagcggtc caccccagac gtggttctgg tcgcccggag gatcaagtat ggaagccgtc
     1861 ataaaggcat ttctgactgg gcaccctgga aaggtgagca agaaggactc caagaagaag
     1921 ccgccagcaa caagcaagaa agacccggag aagacgccct tgctgccaac gagagttaat
     1981 tatattattg gtgtcctggt cttgtgtgag gtaacggggg taagagctga tgtccactta
     2041 ctcgagcagc cagggaacct ttggattaca tgggccaacc gtacaggcca aacggatttt
     2101 tgcctctcta cacagtcagc cacctcccct tttcaaacat gtttgatagg tatcccgtcc
     2161 cctatttccg agggtgattt taagggatat gtttctgata caaattgcac caccttggga
     2221 actgatcggt tagtctcgtc agccgacttt actggcggac ctgacaacag taccaccctc
     2281 acttatcgga aggtctcatg cttgttgtta aagctgaatg tctctatgtg ggatgagcca
     2341 cctgaactac agctgttagg ttcccagtct ctccctaaca ttactaatat tgctcagatt
     2401 tccggtataa ccgggggatg cgtaggcttc agaccacaag gggttccttg gtatctaggt
     2461 tggtctagac aggaggccac gcggtttctc cttagacacc cctctttctc taaatccacg
     2521 gaaccgttta cggtggtgac agcggatagg cacaatcttt ttatggggag tgagtactgc
     2581 ggtgcatatg gctacagatt ttggaacatg tataactgct cacaggtggg gcggcagtac
     2641 cgctgtggta atgcgcgcag cccccgcccg ggtcttcctg aaatccagtg tacaaggaga
     2701 ggaggcaaat gggttaatca atcacaggaa attaatgagt cggagccgtt cagctttacg
     2761 gtgaactgta cagctagtag tttgggtaat gccagtgggt gttgcggaaa agcaggcacg
     2821 attctcccgg gaaagtgggt cgacagcaca caaggtagtt tcaccaaacc aaaagcgcta
     2881 ccacccgcaa ttttcctcat ttgtggggat cgcgcatggc aaggaattcc cagtcgtccg
     2941 gtagggggcc cctgctattt aggcaagctt accatgttag cacctaagca tacagatatt
     3001 ctcaaggtgc ttgtcaattc atcgcggaca ggtataagac gtaaacgaag cacctcacac
     3061 ctggatgata catgctcaga tgaagtgcag ctttggggtc ctacagcaag aatctttgca
     3121 tctatcctag ccccgggggt agcagctgcg caagccttaa gagaaattga gagactagcc
     3181 tgttggtccg ttaaacaggc taacttgaca acatcactcc tcggggactt attggatgat
     3241 gtcacgagta ttcgacacgc ggtcctgcag aaccgagcgg ctattgactt cttgctccta
     3301 gctcacggcc atggctgtga ggacgttgcc ggaatgtgct gtttcaattt gagtgatcac
     3361 agtgagtcta tacagaagaa gttccagcta atgaaggaac atgtcaataa gatcggcgtg
     3421 gacagcgacc caattggaag ttggctgcga ggactattcg ggggaatagg agaatgggcc
     3481 gttcatttgc tgaaaggact gcttttgggg cttgtagtta ttttgttgct agtagtgtgc
     3541 ctgccttgcc ttttgctcga gcgagttggt atttatcttt gcattaaatt aaagcacacc
     3601 aagaaaagac agatttatac agacatagag atgaaccgac ttggaaggta acggatcctt
     3661 cgaaaccggt gctagcgcgc ccctgcaggg aattcgatat caagcttatc gataatcaac
     3721 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3781 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3841 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3901 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3961 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4021 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4081 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     4141 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4201 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4261 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     4321 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     4381 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     4441 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     4501 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     4561 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     4621 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     4681 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     4741 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     4801 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgcc ccggggagct
     4861 caattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     4921 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     4981 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     5041 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     5101 catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga
     5161 acaaaggtgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     5221 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     5281 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     5341 ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca
     5401 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5581 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5641 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5701 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5761 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     5821 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5881 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     5941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6361 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6601 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
