LOCUS       sequence-241781-        8464 bp    DNA     circular SYN 15-APR-2025
DEFINITION  KLRC1-V5 tagged expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8464)
  TITLE     ORFs that regulate PDL1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8464)
  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            "Unpublished"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-KLRC1-V5
FEATURES             Location/Qualifiers
     source          1..8464
                     /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(3325..3349)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3402..3443
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3467..4055
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3520..3540)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3938..3949)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4070..4386
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4237..4372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4434..4481
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4500..4898
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4985..5218
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5290..5411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5327..5346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5381..5400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5451..5586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5606..5625)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5607..5625)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5635..5652)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5635..5651)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5644..5666)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5793..6248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5880..5899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6090..6111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6274..6378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6379..6447,6448..7239)
                     /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(6597..6616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7410..7998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7899..7918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8152..8169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8286..8307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8322..8327,8328..8345,8346..8352)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8360..8376
                     /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     8365..8387
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8384..8400
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8384..8400
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8419..8439
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8421..8439
                     /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 ccatggataa ccaaggagta atctactcag
     2641 acctgaatct gcccccaaac ccaaagaggc agcaacgaaa acctaaaggc aataaaagct
     2701 ccattttagc aactgaacag gaaataacct atgcggaatt aaaccttcaa aaagcttctc
     2761 aggattttca agggaatgac aaaacctatc actgcaaaga tttaccatca gctccagaga
     2821 agctcattgt tgggatcctg ggaattatct gtcttatctt aatggcctct gtggtaacga
     2881 tagttgttat tccctctaca ttaatacaga ggcacaacaa ttcttccctg aatacaagaa
     2941 ctcagaaagc acgtcattgt ggccattgtc ctgaggagtg gattacatat tccaacagtt
     3001 gttactacat tggtaaggaa agaagaactt gggaagagag tttgctggcc tgtacttcga
     3061 agaactccag tctgctttct atagataatg aagaagaaat gaaatttctg tccatcattt
     3121 caccatcctc atggattggt gtgtttcgta acagcagtca tcatccatgg gtgacaatga
     3181 atggtttggc tttcaaacat gagataaaag actcagataa tgctgaactt aactgtgcag
     3241 tgctacaagt aaatcgactt aaatcagccc agtgtggatc ttcaataata tatcattgta
     3301 agcataagct taagggcgaa ttcgacccag ctttcttgta caaagtggtt gatatccagc
     3361 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     3421 ctctcctcgg tctcgattct acgcgtaccg gttagtaatg atcgacaatc aacctctgga
     3481 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3541 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3601 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3661 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3721 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3781 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3841 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3901 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3961 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4021 gacgagtcgg atctcccttt gggccgcctc cccgcctggc gatggtaccg gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4261 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4321 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4381 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     4441 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     4501 tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca
     4561 acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc
     4621 gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg
     4681 tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa
     4741 atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc
     4801 atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc
     4861 gtgaattgct gccctctggt tatgtgtggg agggctaagc acaattcgag ctcggtacct
     4921 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     4981 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     5041 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     5101 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     5161 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     5221 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     5281 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5401 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     5461 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5521 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5581 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     5641 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5701 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5761 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     5821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5941 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6001 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6121 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     6241 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6301 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6361 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6421 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6481 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6541 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6601 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6661 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6721 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6781 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6841 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6901 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6961 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7021 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7081 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7141 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7201 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7261 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8041 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8101 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8161 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     8221 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     8281 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     8341 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     8401 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
     8461 gctt
//
