LOCUS       sequence-241765-        8389 bp    DNA     circular SYN 15-APR-2025
DEFINITION  FGF6-V5 tagged expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8389)
  TITLE     ORFs that regulate PDL1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8389)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLenti6.3-FGF6-V5
FEATURES             Location/Qualifiers
     source          1..8389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1944..2247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2248..2451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2402..2422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2448..2472
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2568..2592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2607..2616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3250..3274)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3327..3368
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3392..3980
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3445..3465)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3863..3874)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3995..4311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4162..4297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4359..4406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4425..4823
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4910..5143
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5215..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5252..5271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5306..5325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5376..5511
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5531..5550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5532..5550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5560..5577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5560..5576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5569..5591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5718..6173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5805..5824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6015..6036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6199..6303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6304..6372,6373..7164)
                     /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(6522..6541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7335..7923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7824..7843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8077..8094
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8211..8232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8247..8252,8253..8270,8271..8277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8285..8301
                     /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     8290..8312
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8309..8325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8309..8325
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8344..8364
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8346..8364
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1981 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2041 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2101 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2161 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2221 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2281 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2341 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2401 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     2461 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
     2521 ctctagagga tccactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca
     2581 aaaaagcagg ctccgaattc gcccttgcca ccatggccct gggacagaaa ctgttcatca
     2641 ctatgtcccg gggagcagga cgtctgcagg gcacgctgtg ggctctcgtc ttcctaggca
     2701 tcctagtggg catggtggtg ccctcgcctg caggcacccg tgccaacaac acgctgctgg
     2761 actcgagggg ctggggcacc ctgctgtcca ggtctcgcgc ggggctagct ggagagattg
     2821 ccggggtgaa ctgggaaagt ggctatttgg tggggatcaa gcggcagcgg aggctctact
     2881 gcaacgtggg catcggcttt cacctccagg tgctccccga cggccggatc agcgggaccc
     2941 acgaggagaa cccctacagc ctgctggaaa tttccaccgt ggagcgaggc gtggtgagtc
     3001 tctttggagt gagaagtgcc ctcttcgtcg ccatgaacag taaaggaaga ttgtacgcaa
     3061 cgcccagctt ccaagaagaa tgcaagttca gagaaaccct cctgcccaac aattacaatg
     3121 cctacgagtc agacttgtac caagggacct acattgccct gagcaaatac ggacgggtaa
     3181 agcggggcag caaggtgtcc ccgatcatga ctgtcactca tttccttccc aggatcaagg
     3241 gcgaattcga cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct
     3301 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     3361 attctacgcg taccggttag taatgatcga caatcaacct ctggattaca aaatttgtga
     3421 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3481 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3541 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3601 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3661 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3721 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3781 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3841 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3901 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3961 cctttgggcc gcctccccgc ctggcgatgg taccggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4141 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4201 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4261 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4321 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     4381 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt
     4441 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     4501 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     4561 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     4621 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     4681 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     4741 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     4801 ctggttatgt gtgggagggc taagcacaat tcgagctcgg tacctttaag accaatgact
     4861 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     4921 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     4981 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5041 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     5101 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     5161 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     5221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5341 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5401 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5461 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     5521 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     5581 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5641 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5701 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6061 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6121 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     6181 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6241 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6301 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6361 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6421 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6481 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6541 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6601 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6661 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6721 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6781 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6841 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6901 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6961 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7021 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7081 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7141 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7201 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7261 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7321 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7381 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7441 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7501 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7561 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7621 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7681 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7741 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7801 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7861 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7921 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7981 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8041 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8101 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8161 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8221 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8281 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8341 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctt
//
