LOCUS       sequence-339916-        7470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivector that expresses LifeAct-Ruby from internal human PGK 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7470)
  TITLE     Lentivirus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7470)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti.PGK.LifeAct-Ruby.W
FEATURES             Location/Qualifiers
     source          1..7470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1972..2482
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2401..2419
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2533..2583
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             complement(2597..2614)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     misc_feature    3321..3909
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3374..3394)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3792..3803)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3996..4229
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4301..4422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4338..4357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4392..4411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4462..4597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4617..4636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4618..4636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4646..4663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4646..4662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4655..4677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4804..5259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4891..4910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5101..5122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5285..5389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5390..5458,5459..6250)
                     /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(5608..5627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6421..7009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6910..6929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7163..7180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7297..7318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7333..7338,7339..7356,7357..7363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7371..7387
                     /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     7376..7398
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7395..7411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7395..7411
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7430..7450
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7432..7450
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag aagcttgata tcgaattcca cggggttggg
     1981 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     2041 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     2101 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     2161 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     2221 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     2281 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     2341 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     2401 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     2461 aatcaccgac ctctctcccc agggggatcc tagcgctacc ggactcagat ctcgagctca
     2521 agcttcgaat tcatgggtgt cgcagatttg atcaagaaat tcgaaagcat ctcaaaggaa
     2581 gaaggggatc cgggtgctgg tgctggtgct ggtgctatgg gcaagcttac catggccagc
     2641 tccgaggatg tcatcaaaga gtttatgaga tttaaggtca agatggaggg aagcgtcaac
     2701 ggacacgagt tcgagattga gggagaagga gaaggccggc cttacgaggg cacacaaacc
     2761 gctaagctca aggtcacaaa aggaggaccc ctccccttct cctgggatat tctgagccct
     2821 cagttccagt acggaagcaa agcctatgtt aaacaccctg ccgacatccc tgactatctg
     2881 aagctctcct tccctgaagg cttcaagtgg gagagattca tgaacttcga ggacggaggc
     2941 gtggtgacag tcacacaaga tagcaccctc caggacggag agtttattta taaggtgaaa
     3001 ctcagaggaa ccaacttccc ctccgatggc cctgtcatgc aaaaaaaaac aatgggatgg
     3061 gaagcctcca ccgagagaat gtatcctgag gatggcgctc tgaaaggcga aattaaaatg
     3121 agactgaaac tcaaagacgg aggacactac gatgccgagg tcaaaacaac ctacaaggcc
     3181 aagaaacaag tgcagctgcc tggcgcctac atgactgata ttaaactcga cattatcagc
     3241 cataatgggg actacaccat cgtggaacaa tatgagagag ctgagggcag acatagcaca
     3301 ggcgcttaag cggcctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3361 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3421 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3481 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3541 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3601 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3661 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3721 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3781 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3841 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3901 cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
     3961 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4021 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     4081 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     4141 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     4201 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     4261 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     4321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     4441 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     4621 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4681 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4741 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4801 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5221 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     5281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5341 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5401 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5461 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5521 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5581 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5641 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5701 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5761 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5941 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6001 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6241 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6541 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6781 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7081 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7141 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     7201 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     7261 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     7321 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     7381 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     7441 tcactaaagg gaacaaaagc tggagctgca
//
