LOCUS       sequence-417653-        7614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codes for Gag from HIV-1 with an internal mCherry fluorescent 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  AUTHORS   Diana Brychka , Nilda Vanesa Ayala-Nunez, Yonis Bare, Amandine 
            Dupas, Emma Partiot, Vincent Mittelheisser, Vincent Lucansky, Jacky 
            G. Goetz, Nael Osmani, Raphael Gaudin
  TITLE     Targeting monocytic Occludin impairs monocyte transmigration and HIV
            neuroinvasion
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 7614)
  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: "bioRxiv"
COMMENT     Sequence Label: pcDNA3.3 Gag-imCherry
FEATURES             Location/Qualifiers
     source          1..7614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(160..264)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(339..358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        530..909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        910..1113
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1064..1084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1110..1134
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1209..1218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1629..2333
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1771..1789)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2039..2058
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3480..3499)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3524..3572
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3774..4202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3861..3880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4071..4092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4211..4231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4216..4545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4458..4477
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4612..5406
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4666..4685)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5276..5295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5582..5703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5619..5638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5673..5692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5752..5768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5752..5768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5765..5787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5776..5792
                     /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(5800..5806,5807..5824,5825..5830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5845..5866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5983..6000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6154..6742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6234..6253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6913..7614,1..90,91..159))
                     /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     7536..7555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
       61 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
      301 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
      361 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
      421 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
      481 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      541 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      601 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      661 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      721 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      781 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      841 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      901 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      961 cggggatttc caagtctcta ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     1021 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     1081 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     1141 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
     1201 actctagcgc caccatgggt gcgagagcgt cggtattaag cgggggagaa ttagataaat
     1261 gggaaaaaat tcggttaagg ccagggggaa agaaacaata taaactaaaa catatagtat
     1321 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg ccttttagag acatcagaag
     1381 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1441 gatcattata taatacaata gcagtcctct attgtgtgca tcaaaggata gatgtaaaag
     1501 acaccaagga agccttagat aagatagagg aagagcaaaa caaaagtaag aaaaaggcac
     1561 agcaagcagc agctgacaca ggaaacaaca gccaggtctc gcagaactat ccaattgtac
     1621 aaacgcgtgt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     1681 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     1741 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1801 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     1861 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1921 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1981 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2041 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2101 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2161 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2221 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2281 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtctagaa
     2341 gccaaaatta ccctatagtg cagaacctcc aggggcaaat ggtacatcag gccatatcac
     2401 ctagaacttt aaatgcatgg gtaaaagtag tagaagagaa ggctttcagc ccagaagtaa
     2461 tacccatgtt ttcagcatta tcagaaggag ccaccccaca agatttaaat accatgctaa
     2521 acacagtggg gggacatcaa gcagccatgc aaatgttaaa agagaccatc aatgaggaag
     2581 ctgcagaatg ggatagattg catccagtgc atgcagggcc tattgcacca ggccagatga
     2641 gagaaccaag gggaagtgac atagcaggaa ctactagtac ccttcaggaa caaataggat
     2701 ggatgacaca taatccacct atcccagtag gagaaatcta taaaagatgg ataatcctgg
     2761 gattaaataa aatagtaaga atgtatagcc ctaccagcat tctggacata agacaaggac
     2821 caaaggaacc ctttagagac tatgtagacc gattctataa aactctaaga gccgagcaag
     2881 cttcacaaga ggtaaaaaat tggatgacag aaaccttgtt ggtccaaaat gcgaacccag
     2941 attgtaagac tattttaaaa gcattgggac caggagcgac actagaagaa atgatgacag
     3001 catgtcaggg agtgggggga cccggccata aagcaagagt tttggctgaa gcaatgagcc
     3061 aagtaacaaa tccagctacc ataatgatac agaaaggcaa ttttaggaac caaagaaaga
     3121 ctgttaagtg tttcaattgt ggcaaagaag ggcacatagc caaaaattgc agggccccta
     3181 ggaaaaaggg ctgttggaaa tgtggaaagg aaggacacca aatgaaagat tgtactgaga
     3241 gacaggctaa ttttttaggg aagatctggc cttcccacaa gggaaggcca gggaattttc
     3301 ttcagagcag accagagcca acagccccac cagaagagag cttcaggttt ggggaagaga
     3361 caacaactcc ctctcagaag caggagccga tagacaagga actgtatcct ttagcttccc
     3421 tcagatcact ctttggcagc gacccctcgt cacaataagt ttaaacgggg gaggctaact
     3481 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3541 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3601 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3661 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3721 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3781 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3841 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3901 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3961 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4021 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4081 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4141 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4201 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4321 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4561 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4621 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4681 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4741 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4801 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4861 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4921 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4981 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5041 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5101 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5161 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5221 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5281 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5341 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5401 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggat
//
