LOCUS       sequence-228221-        7204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a 3rd generation lentiviral transfer vector (slightly lower 
            titer, highest expression level) from Nolan lab's FELIX vector 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7204)
  TITLE     Nolan Lab Plasmids
REFERENCE   2  (bases 1 to 7204)
  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"
COMMENT     Sequence Label: pLionII
FEATURES             Location/Qualifiers
     source          1..7204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(288..307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     407..429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(467..485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        553..657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(658..726,727..1518)
                     /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(876..895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1689..2277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2431..2448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2565..2586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2601..2606,2607..2624,2625..2631)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2639..2655
                     /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     2644..2666
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2663..2679
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2663..2679
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2970..2986
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2971..2987
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2990..3578
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3096..3107
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3505..3525
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     promoter        complement(4436..4454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4499..4702)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        4703..5082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(5128..5146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6206..6409)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        6410..6789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
       61 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
      121 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
      181 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
      241 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
      301 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      361 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      421 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
      481 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
      541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      601 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      661 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      721 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      781 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      841 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      901 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      961 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1021 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1081 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1141 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1201 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1261 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1321 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1381 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1441 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1501 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2341 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2401 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2461 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     2521 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2581 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2641 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
     2701 tgcctgcagt gcgaagttct cggcccggat tccgagacct cacaggtaaa aaagattaca
     2761 cagatactcg acagggttca atctcaaata tttattgtat ctgtgggagc ctcaagggag
     2821 aactcaaaag tcctcaacaa agagactcct cgaagtttca caaagcactg gttacctagg
     2881 agctgatcaa tcaggccttc agggttccaa tactcatccc agtccaccct tttttctttt
     2941 ttaggaggtt catcttcagg ggccccccct cgaggtcgac ggtatcgatg cggggaggcg
     3001 gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga
     3061 gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag
     3121 aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat
     3181 gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag
     3241 cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc
     3301 ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac
     3361 acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc
     3421 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     3481 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     3541 gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt gattgaaggc
     3601 tttcttcttt cttcttcttc tttgtctttt ccttttacct tacaatacat actttattag
     3661 tttgatttga ttcgaaatgg attcatatga cataccttcc tcaaagggaa gaactcagca
     3721 cagtatctcc cttataaaat aagtctgaga tacttcatca ttcctcctct ttttcagaca
     3781 tgccacattg cctaccattt ctcctcaatt ccatttgtgg ttgtatttct tctccttcta
     3841 cttcaggcat tgcaattact gtgtatccta gtatcttgtg gatacaattt cttatacaat
     3901 caaccaatgt aggtaaacat aaaatcaata ataacactcc taatcctatt cccaagatac
     3961 ctcccaatag tccctttaaa tattgtggaa tatttcctat ccatcttacc caatcttccc
     4021 acttttgtaa ttgttgtatc cctgttttcc cttgtacatt attttgttct atgtccatta
     4081 ttatttcata aaacttttgt tgtaaatctt ttgtttggtt ataccattcc cccaaagtta
     4141 tatttccatg attccatatt gtttgattta tagtcatatt ataccttgtc cacaactcaa
     4201 gagggatttt gcagaagaat tgattttgat tacatcctaa ttcttgcata gcgaaagctg
     4261 tatataaaaa tttttccata gcttctactt ttaatcctat tactagcttc tattaggcgc
     4321 gctagactcg agcggccgcc actgtgctgg atatctgcag aattccacca cactggacta
     4381 gtggatccga gctcggtacc aagcttaagt ttaaacgcta gccagcttgg gtctccctat
     4441 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     4501 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     4561 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     4621 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     4681 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4741 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4801 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4861 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4921 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4981 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     5041 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc
     5101 gtacatcggg cccggtaccc aattcgccct atagtgagtc gtattacgcg cgccgtcata
     5161 tttaaaagtc ctgctaccgc aaagactttt aatgtcacaa tttacatgtg gaatcaggat
     5221 atatatgggc acaaataatt tctcaagaaa ctgctgactg tacagttaaa gctgttttac
     5281 aattgttgag tgctcataat gttactgaat tacaaacaga taatggacca aattttaaaa
     5341 atcaaaagat ggaaggagtt acccacaaga attattttgc catcaaagtg tgtgcaatcc
     5401 atttgccaga taccaggccc tattttcaat tgtcctccca cttgttctcc tctgattctg
     5461 catactgggc attttcgtct tatctcttct gctaccatct taggtaaatt aaattcagtc
     5521 cttaagtaat tgccatatca atttctttgc tagatccaaa tttttctctt ctttcttgta
     5581 aatcgcaaat aaccaacctt agttgatcta aagtctctgg tatatcacca ggttctcgtc
     5641 ctgtagatac attagccatt ctaatggccc atctgaaatt cccttctcca aattttttac
     5701 tcttcccccc tactcctaca gcaacattac tacatctctt aatggccatt ttccaatctc
     5761 gcccctgtcc attccccatg ttgctgtaga atctctccta ccttgttgac tgtccctcgg
     5821 cgaatctcct ggcttgaagg tccgcgaaga gtcaccagat gtaatttatc tgggccttta
     5881 aacaatgact tgattatgag ctcgagtctg cttcactaga gatactcact gttagcagcg
     5941 tctgctactg cttccctatt taggtcagca ggagttctgc ttaacagctt tctattgctc
     6001 tagcttcact tcctcaatca ctctcaatca agtccctgtt cgggcgccaa ctgcgaagtt
     6061 ctcggcccgg attccgagac ctcacaggta aaaaagatta cacagatact cgacagggtt
     6121 caatctcaaa tatttattgt atctgtggga gcctcaaggg agaactcaaa agtcctcaac
     6181 aaagagactc ctcgaagttt cacagagctc tgcttatata gacctcccac cgtacacgcc
     6241 taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag tcccgttgat
     6301 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     6361 agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc atggtaatag
     6421 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc atgtactggg
     6481 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga cttggcatat
     6541 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     6601 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     6661 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     6721 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     6781 atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac atattatgat
     6841 atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga
     6901 ataatatggc taatggccaa tattgaagat ctgtcgagcc atgtgagcaa aaggccagca
     6961 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7021 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7081 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7141 gcttaccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7201 tgct
//
