LOCUS       sequence-393119-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral transfer plasmid for constitutive expression of a GFP in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  AUTHORS   Stevens TA, Tomaleri GP, Hazu M, Wei S, Nguyen VN, DeKalb C, 
            Voorhees RM, Pleiner T
  TITLE     A nanobody-based strategy for rapid and scalable purification of 
            human protein complexes.
  JOURNAL   Nat Protoc. 2024 Jan;19(1):127-158. doi: 10.1038/s41596-023-00904-w.
            Epub 2023 Nov 16.
  PUBMED    37974029
REFERENCE   2  (bases 1 to 7094)
  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: 
            "10.1038/s41596-023-00904-w"; journalName: "Nat Protoc"; date: 
            "2024-01"; volume: "19"; issue: "1"; pages: "127-158"
COMMENT     Sequence Label: pTP924 pHAGE2 CMV GFP
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             20..653
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    700..825
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1322..1555
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1740..1784
                     /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"
     misc_feature    2077..2194
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2246..2549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2550..2753
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2704..2724
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2750..2774
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2846..2855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3555..4143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3608..3628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4026..4037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4291..4471
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4518..4536)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4604..4708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4709..4777,4778..5569)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4927..4946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5740..6328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6739..6760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6775..6780,6781..6798,6799..6805)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6813..6829
                     /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     6818..6840
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6837..6853
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6837..6853
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6903..6922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtactacaaa cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt
       61 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
      121 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
      181 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
      241 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
      301 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
      361 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
      421 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
      481 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      541 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      601 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      661 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      721 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      781 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      841 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      901 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      961 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     1021 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     1081 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     1141 agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata
     1201 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1261 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1321 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1381 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1441 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1501 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1561 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1621 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     1681 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     1741 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     1801 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     1861 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     1921 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     1981 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
     2041 atcgccgaat tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     2101 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2161 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     2221 tccagtttgg actagtggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2281 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2341 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2401 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2461 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2521 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2581 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2641 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2701 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     2761 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
     2821 ggctagtgtt taaacttaag ctgcggccgc catggtgtct aaaggtgaag aactttttac
     2881 gggtgtcgtc cccatcctgg ttgaattgga cggagacgtg aatggccata aattttcagt
     2941 gagcggtgaa ggagagggag acgccactta cggtaaactt accttgaaat ttatctgcac
     3001 taccgggaaa ctgcccgtac cctggccgac gctcgtaaca acgcttacct acggcgtgca
     3061 atgttttagc agatacccag accatatgaa gcagcacgat tttttcaaga gtgcaatgcc
     3121 tgaggggtac gtacaggaaa ggacgatctt tttcaaagac gatggtaatt ataaaacgcg
     3181 ggctgaggtt aaatttgaag gagacacgct cgtgaatcga atagagctga agggaatcga
     3241 cttcaaggaa gacggtaaca tcctgggcca taaactcgaa tataattaca attcacacaa
     3301 cgtctatatc atggcagata aacagaagaa tggcatcaaa gtcaatttta agatccggca
     3361 taatatcgag gacggatcag tacagcttgc cgaccattac caacagaata caccgatagg
     3421 agacggtccc gtcctcttgc cagataatca ctatttgagc acacaatccg ctctttctaa
     3481 agatccgaat gaaaagcgcg atcatatggt cttgaaagag ttcgtcactg ccgccggcat
     3541 aacctgactc gagtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3601 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3661 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3721 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3781 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3841 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3901 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gttggggaaa tcatcgtcct
     3961 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4021 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4081 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4141 cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     4201 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tcacctgcag
     4261 gacaggcgcg ccctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     4321 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     4381 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     4441 cccttttagt cagtgtggaa aatctctagc acccgggcga ttaaggaaag ggctagatca
     4501 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4561 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4681 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4741 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4801 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4861 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4921 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4981 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5041 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5101 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5161 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5221 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5281 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5341 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5401 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5461 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5521 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5581 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5641 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5761 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5821 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5881 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5941 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6001 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6061 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6121 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6181 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6241 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6301 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6361 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6421 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6481 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6541 gcgttggccg attcattaat gcagcaagct catggctgac taattttttt tatttatgca
     6601 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6661 ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact ggaaagcggg
     6721 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6781 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6841 aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac tgcattctag
     6901 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca
     6961 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct
     7021 gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg
     7081 tatttgttaa atat
//
