LOCUS       sequence-337165-        7298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expressing HIV-1 envelope glycoprotein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7298)
  AUTHORS   Hamilton JR, Tsuchida CA, Nguyen DN, Shy BR, McGarrigle ER, Sandoval
            Espinoza CR, Carr D, Blaeschke F, Marson A, Doudna JA
  TITLE     Targeted delivery of CRISPR-Cas9 and transgenes enables complex 
            immune cell engineering.
  JOURNAL   Cell Rep. 2021 Jun 1;35(9):109207. doi: 
            10.1016/j.celrep.2021.109207.
  PUBMED    34077734
REFERENCE   2  (bases 1 to 7298)
  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"; journalName: "Cell Rep.";
            date: "2021-06-1"; volume: "35(9):109207. doi"; pages: " 
            10.1016/j.celrep.2021.109207"
COMMENT     Sequence Label: pCAT001
FEATURES             Location/Qualifiers
     source          1..7298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3702..3746
                     /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"
     primer_bind     complement(4320..4339)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4385..4440
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4439..4458)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4801..4817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4801..4817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4814..4836)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4825..4841
                     /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(4849..4855,4856..4873,4874..4879))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4894..4915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4974..5169
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5020..5155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5175..5309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5212..5231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5266..5285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5377..5394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5628..5647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6307..7098,7099..7167))
                     /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     6930..6949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7168..7272)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcgccacca
     1741 tgagagtcaa ggaaaagtac caacacttgt ggcgatgggg gtggaggtgg ggaaccatgt
     1801 tgttgggaat gttgatgatc tgctcagcaa cagaaaagtt gtgggtgact gtttattatg
     1861 gcgtgcctgt atggaaggaa gcgactacaa cccttttttg cgcttccgat gctaaagcct
     1921 acgatacgga agtccacaat gtttgggcca cgcacgcatg cgtaccaaca gacccgaacc
     1981 cgcaggaagt cgtcctcgtg aatgtgacgg agaactttaa tatgtggaag aatgacatgg
     2041 ttgagcagat gcacgaggac attatttcac tttgggacca gtcccttaaa ccttgcgtga
     2101 agcttacacc actgtgtgtc agcctcaagt gcactgacct gaaaaatgac acaaatacca
     2161 actcaagctc tggtcgcatg atcatggaaa aaggtgagat taaaaattgt tcttttaata
     2221 taagcacctc cattcgggga aaggtacaaa aagaatacgc ctttttctat aaacttgaca
     2281 tcatcccgat tgataacgac actactagtt ataaacttac ctcctgcaat acgagcgtca
     2341 tcacacaggc ttgcccgaag gtgtctttcg agccgattcc catacattac tgtgcaccgg
     2401 ccggttttgc catcctgaaa tgcaataata aaacgtttaa cggcacgggg ccttgcacga
     2461 acgtaagcac tgtacaatgc actcatggga taagacctgt ggtgtctact cagctccttc
     2521 tgaacggatc actcgccgag gaggaagtag tgatacggtc agtaaacttt accgacaacg
     2581 ccaagactat catagttcag cttaacacga gcgtcgagat caattgcact cggcccaata
     2641 ataatactcg caaaaggatt cgcatccaac gcgggcccgg tcgagcgttc gtgacgattg
     2701 ggaagatagg caatatgcgc caggcgcatt gtaacatatc tcgcgctaag tggaataata
     2761 ccttgaaaca gatagctagc aagctccgag aacaatttgg gaacaataaa actataatat
     2821 ttaagcaatc ttccgggggt gatcctgaaa ttgtcactca ctcttttaat tgtggcggag
     2881 agttctttta ttgcaactca acccagctgt tcaattccac ttggtttaat agcacgtggt
     2941 caacagaagg ttctaacaac acagagggca gcgacacaat tacactgcca tgtcggatca
     3001 aacaaatcat caacatgtgg caaaaggttg gcaaggcgat gtatgccccg ccgatttcag
     3061 gccagataag gtgcagttct aatatcacgg gccttttgtt gactagagac ggagggaact
     3121 ccaacaacga gagcgaaatt tttcgacctg gaggggggga tatgcgcgat aactggagga
     3181 gcgagttgta caaatacaaa gtggtcaaga tagagcctct cggagtagcg ccaaccaaag
     3241 ctaaacgccg cgttgtccag cgcgagaaac gggccgtggg cataggtgct ctctttcttg
     3301 ggttcctcgg ggcagccgga tccactatgg gtgccgcgtc aatgaccttg acggtccagg
     3361 ccagacaact ccttagtggc atcgtccagc agcagaataa ccttctgcgg gccatcgaag
     3421 cccaacagca ccttctccag ctcaccgttt gggggataaa acaactccaa gcgcgcatac
     3481 ttgccgtaga gcgatacctc aaggatcaac aattgcttgg tatctggggc tgttcaggaa
     3541 aacttatctg tacaacggca gtgccatgga acgcatcttg gagtaacaaa agcctggagc
     3601 agatatggaa ccatactacg tggatggaat gggataggga gatcaataat tacacgagcc
     3661 ttatccattc tttgatcgag gagagtcaga accagcaaga gaagaatgag caagaattgc
     3721 tggagttgga taaatgggct tccctctgga actggttcaa tattacgaac tggttgtggt
     3781 acattaagct cttcattatg atcgtagggg ggctggtagg cctgcggatt gtgtttgcgg
     3841 tacttagtat cgtgaatcga gtacgacaag ggtattcccc gctgtctttc cagacacact
     3901 tgcctacacc gaggggccca gaccgaccgg agggtatcga agaagaaggg ggcgagcgcg
     3961 atagggacag gagtatccgc cttgtaaacg gctccttggc cctcatttgg gacgatctta
     4021 gatcactttg tctcttttcc tatcatcggc ttagggattt gcttctgatc gtgacgagga
     4081 tcgttgagct cttgggtagg cgaggctggg aggcactgaa gtattggtgg aaccttctcc
     4141 aatactggtc ccaagaactc aagaactccg ctgtcagctt gttgaacgca acagccattg
     4201 ctgtcgctga gggaacagac cgggtgatcg aagtagtgca aggggcttgt cgagcgataa
     4261 ggcacattcc ccgacgcatt cgccaaggac tggagagaat tctgctctag gctagcagat
     4321 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     4381 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     4441 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     4501 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     4561 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     4621 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     4681 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     4741 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     4801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     4981 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5041 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5101 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5161 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5221 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5281 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7261 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
