LOCUS       sequence-403501-        7655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for C terminal mCherry fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7655)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 7655)
  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"
COMMENT     Sequence Label: pLenti6-V5-Topo mCherry
FEATURES             Location/Qualifiers
     source          1..7655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             627..1337
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(772..790)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1040..1059
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1350..1391
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1428..1448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1433..1762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1613..1748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1810..1857
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1876..2274
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2361..2594
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2666..2787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2757..2776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2827..2962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2982..3001)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2983..3001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3011..3028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3011..3027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3020..3042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3169..3624
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3256..3275)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3466..3487
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3650..3754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3755..3823,3824..4615)
                     /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     complement(3973..3992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4786..5374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5275..5294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5528..5545
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5662..5683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5698..5703,5704..5721,5722..5728)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5736..5752
                     /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     5741..5763
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5760..5776
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5760..5776
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5795..5815
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5797..5815
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5843..6069
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6070..6250
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6297..6422
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6915..7148
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7333..7377
                     /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"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagaggatcc acgctgcgtc
      601 tgtttaaacg ccacgagcat ctcgagatgg tgagcaaggg cgaggaggat aacatggcca
      661 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      721 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      781 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      841 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      901 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      961 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1021 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1081 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1141 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1201 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1261 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1321 acgagctgta caagtagccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     1381 tcgattctac gcgtaccggt tagtaatgag tttggaatta attctgtgga atgtgtgtca
     1441 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1501 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1561 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1621 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1681 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     1741 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     1801 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     1861 gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     1921 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     1981 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     2041 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     2101 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     2161 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     2221 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcacaa
     2281 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     2341 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     2401 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2461 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2521 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2581 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     2641 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     2701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2761 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     2821 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2881 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2941 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     3001 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3061 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3121 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     3181 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3241 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3301 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3361 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3421 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3481 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3541 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3601 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3661 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3721 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3901 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3961 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4021 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4081 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4141 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4201 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4261 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4321 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4381 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4441 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4501 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4561 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4621 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4681 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4741 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4801 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4861 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4921 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4981 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5041 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5101 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5161 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5221 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5281 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5341 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5401 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5461 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5521 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5641 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5701 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5761 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     5821 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     5881 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
     5941 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
     6001 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
     6061 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6121 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6181 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6241 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     6301 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     6361 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     6421 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     6481 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     6541 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     6601 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     6661 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     6721 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     6781 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     6841 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     6901 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     6961 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     7021 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     7081 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     7141 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     7201 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     7261 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     7321 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     7381 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     7441 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     7501 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     7561 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     7621 ggatctcgac ggtatcgata agcttgggag ttccg
//
