LOCUS       sequence-235249-        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for matrix domain of HIV-1 Gag fused to
            the N-terminal half of split fluorescent Venus (VN).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  AUTHORS   Heredia JD, Park J, Choi H, Gill KS, Procko E
  TITLE     Conformational engineering of HIV-1 Env based on mutational 
            tolerance in the CD4 and PG16 bound states.
  JOURNAL   J Virol. 2019 Mar 20. pii: JVI.00219-19. doi: 10.1128/JVI.00219-19.
  PUBMED    30894475
REFERENCE   2  (bases 1 to 4866)
  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: "J Virol. 
            2019 Mar 20. pii: JVI.00219-19. doi: 10.1128/JVI.00219-19."
COMMENT     Sequence Label: pCMV-MA-VN
FEATURES             Location/Qualifiers
     source          1..4866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(314..333)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    358..405
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      734..1322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1223..1242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1497..1800
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1801..2004
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1955..1975
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2501..2503,2504..2962)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(2543..2564)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2804..2823)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3090..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3127..3146)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3181..3200
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3218..3673)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3355..3376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3567..3586
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3700..3804
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3806..4163
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3806..3997
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3829..3849)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4014..4149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4198..4866,1..126)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(4198..4866,1..126)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4252..4271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4862..4866,1..15)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
       61 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
      121 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
      181 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      241 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
      301 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
      361 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
      421 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
      481 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
      541 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
      601 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      661 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      721 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      781 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      841 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      901 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      961 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1021 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1081 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1141 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1201 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1261 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1321 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1381 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     1441 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1501 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1561 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1621 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1681 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1741 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1801 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1861 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1921 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1981 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     2041 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccggac cgatccagcc
     2101 tccgcgggcg cgcgtcgaca gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     2161 attagatcga tgggaaaaaa ttcggttaag gccaggggga aagaagaagt acaagctaaa
     2221 gcacatcgta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     2281 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     2341 agaggagctt cgatcactat acaacacagt agcaaccctc tattgtgtgc accagcggat
     2401 cgagatcaag gacaccaagg aagctttaga caagatagag gaagagcaaa acaagtccaa
     2461 gaagaaggcc cagcaggcag cagctgacac aggacacagc gtgagcaagg gcgaggagct
     2521 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2581 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     2641 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     2701 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2761 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2821 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2881 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2941 ccacaacgtc tatctgaccg cctaatgagc ggccgcgact ctagatcata atcagccata
     3001 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3061 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3121 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3181 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3241 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3301 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3361 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3421 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3481 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3541 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3601 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3661 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3781 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3841 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3961 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4081 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4141 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4861 gcgttg
//
