LOCUS       sequence-241746-        8437 bp    DNA     circular SYN 15-APR-2025
DEFINITION  CD300C-V5 tagged expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8437)
  TITLE     ORFs that regulate PDL1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8437)
  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            "Unpublished"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-CD300C-V5
FEATURES             Location/Qualifiers
     source          1..8437
                     /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"
     protein_bind    complement(3298..3322)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3375..3416
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3440..4028
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3493..3513)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3911..3922)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4043..4359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4210..4345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4407..4454
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4473..4871
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4958..5191
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5263..5384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5300..5319)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5354..5373
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5424..5559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5579..5598)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5580..5598)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5608..5625)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5608..5624)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5617..5639)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5766..6221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5853..5872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6063..6084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6247..6351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6352..6420,6421..7212)
                     /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(6570..6589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7383..7971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7872..7891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8125..8142
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8259..8280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8295..8300,8301..8318,8319..8325)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8333..8349
                     /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     8338..8360
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8357..8373
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8357..8373
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8392..8412
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8394..8412
                     /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 ccatgactgc cagggcctgg gcctcgtggc
     2641 ggtcttcagc tctgctcctc ctgcttgtcc caggctattt tcctctgagc caccccatga
     2701 ccgtggcggg ccccgtgggg ggatccctga gtgtgcagtg tcgctatgag aaggaacaca
     2761 ggaccctcaa caaattctgg tgcagaccac cacagattct ccgatgtgac aagattgtgg
     2821 agaccaaagg gtcagcaggg aaaaggaatg gccgagtgtc catcagggac agtcctgcaa
     2881 acctcagctt cacagtgacc ctggagaatc tcacagagga ggacgcaggc acctactggt
     2941 gtggggtgga tacaccgtgg ctccgagact ttcatgatcc cattgtcgag gttgaggtgt
     3001 ccgtgttccc ggccgggacg accacagcct ccagccccca gagctccatg ggcacctcag
     3061 gtcctcccac gaagctgccc gtgcacacct ggcccagcgt gaccagaaag gacagccccg
     3121 aacccagccc acaccctggc tccctgttca gcaatgtccg cttcctgctc ctggtcctct
     3181 tggagctgcc cctgctcctg agcatgctgg gcgccgtcct ctgggtgaac agacctcaga
     3241 gaagctctag aagcaggcag aattggccca agggtgagaa ccagaagggc gaattcgacc
     3301 cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg agtctagagg
     3361 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     3421 ccggttagta atgatcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3481 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3541 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3601 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3661 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3721 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3781 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3841 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3901 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3961 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4021 ctccccgcct ggcgatggta ccggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4381 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     4441 tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat
     4501 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     4561 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     4621 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     4681 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     4741 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     4801 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     4861 gggagggcta agcacaattc gagctcggta cctttaagac caatgactta caaggcagct
     4921 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     4981 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5041 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5101 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5161 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     5221 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     5281 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5341 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     5401 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5461 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5521 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     5581 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5641 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5701 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5761 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5821 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5881 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5941 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6001 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6061 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6121 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6181 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     6241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8041 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8101 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     8161 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8221 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8281 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8341 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     8401 cctcactaaa gggaacaaaa gctggagctg caagctt
//
