LOCUS       sequence-332724-        7289 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 7289)
  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 7289)
  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..7289
                     /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"
     CDS             3693..3737
                     /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(4311..4330)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4376..4431
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4430..4449)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4792..4808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4792..4808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4805..4827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4816..4832
                     /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(4840..4846,4847..4864,4865..4870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4885..4906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4965..5160
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5011..5146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5166..5300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5203..5222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5257..5276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5368..5385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5539..6127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5619..5638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6298..7089,7090..7158))
                     /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     6921..6940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7159..7263)
                     /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 attcgccacc atgagagtca
     1741 aggaaaagta ccaacacttg tggcgatggg ggtggaggtg gggaaccatg ttgttgggaa
     1801 tgttgatgat ctgctcagca acagaaaagt tgtgggtgac tgtttattat ggcgtgcctg
     1861 tatggaagga agcgactaca accctttttt gcgcttccga tgctaaagcc tacgatacgg
     1921 aagtccacaa tgtttgggcc acgcacgcat gcgtaccaac agacccgaac ccgcaggaag
     1981 tcgtcctcgt gaatgtgacg gagaacttta atatgtggaa gaatgacatg gttgagcaga
     2041 tgcacgagga cattatttca ctttgggacc agtcccttaa accttgcgtg aagcttacac
     2101 cactgtgtgt cagcctcaag tgcactgacc tgaaaaatga cacaaatacc aactcaagct
     2161 ctggtcgcat gatcatggaa aaaggtgaga ttaaaaattg ttcttttaat ataagcacct
     2221 ccattcgggg aaaggtacaa aaagaatacg cctttttcta taaacttgac atcatcccga
     2281 ttgataacga cactactagt tataaactta cctcctgcaa tacgagcgtc atcacacagg
     2341 cttgcccgaa ggtgtctttc gagccgattc ccatacatta ctgtgcaccg gccggttttg
     2401 ccatcctgaa atgcaataat aaaacgttta acggcacggg gccttgcacg aacgtaagca
     2461 ctgtacaatg cactcatggg ataagacctg tggtgtctac tcagctcctt ctgaacggat
     2521 cactcgccga ggaggaagta gtgatacggt cagtaaactt taccgacaac gccaagacta
     2581 tcatagttca gcttaacacg agcgtcgaga tcaattgcac tcggcccaat aataatactc
     2641 gcaaaaggat tcgcatccaa cgcgggcccg gtcgagcgtt cgtgacgatt gggaagatag
     2701 gcaatatgcg ccaggcgcat tgtaacatat ctcgcgctaa gtggaataat accttgaaac
     2761 agatagctag caagctccga gaacaatttg ggaacaataa aactataata tttaagcaat
     2821 cttccggggg tgatcctgaa attgtcactc actcttttaa ttgtggcgga gagttctttt
     2881 attgcaactc aacccagctg ttcaattcca cttggtttaa tagcacgtgg tcaacagaag
     2941 gttctaacaa cacagagggc agcgacacaa ttacactgcc atgtcggatc aaacaaatca
     3001 tcaacatgtg gcaaaaggtt ggcaaggcga tgtatgcccc gccgatttca ggccagataa
     3061 ggtgcagttc taatatcacg ggccttttgt tgactagaga cggagggaac tccaacaacg
     3121 agagcgaaat ttttcgacct ggaggggggg atatgcgcga taactggagg agcgagttgt
     3181 acaaatacaa agtggtcaag atagagcctc tcggagtagc gccaaccaaa gctaaacgcc
     3241 gcgttgtcca gcgcgagaaa cgggccgtgg gcataggtgc tctctttctt gggttcctcg
     3301 gggcagccgg atccactatg ggtgccgcgt caatgacctt gacggtccag gccagacaac
     3361 tccttagtgg catcgtccag cagcagaata accttctgcg ggccatcgaa gcccaacagc
     3421 accttctcca gctcaccgtt tgggggataa aacaactcca agcgcgcata cttgccgtag
     3481 agcgatacct caaggatcaa caattgcttg gtatctgggg ctgttcagga aaacttatct
     3541 gtacaacggc agtgccatgg aacgcatctt ggagtaacaa aagcctggag cagatatgga
     3601 accatactac gtggatggaa tgggataggg agatcaataa ttacacgagc cttatccatt
     3661 ctttgatcga ggagagtcag aaccagcaag agaagaatga gcaagaattg ctggagttgg
     3721 ataaatgggc ttccctctgg aactggttca atattacgaa ctggttgtgg tacattaagc
     3781 tcttcattat gatcgtaggg gggctggtag gcctgcggat tgtgtttgcg gtacttagta
     3841 tcgtgaatcg agtacgacaa gggtattccc cgctgtcttt ccagacacac ttgcctacac
     3901 cgaggggccc agaccgaccg gagggtatcg aagaagaagg gggcgagcgc gatagggaca
     3961 ggagtatccg ccttgtaaac ggctccttgg ccctcatttg ggacgatctt agatcacttt
     4021 gtctcttttc ctatcatcgg cttagggatt tgcttctgat cgtgacgagg atcgttgagc
     4081 tcttgggtag gcgaggctgg gaggcactga agtattggtg gaaccttctc caatactggt
     4141 cccaagaact caagaactcc gctgtcagct tgttgaacgc aacagccatt gctgtcgctg
     4201 agggaacaga ccgggtgatc gaagtagtgc aaggggcttg tcgagcgata aggcacattc
     4261 cccgacgcat tcgccaagga ctggagagaa ttctgctcta ggctagcaga tctttttccc
     4321 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4381 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4441 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4501 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     4561 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     4621 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     4681 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4741 cttctcttat gaagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     4801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     4981 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5041 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5101 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5161 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5221 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5281 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5701 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5941 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7261 gcgcacattt ccccgaaaag tgccacctg
//
