LOCUS       sequence-236040-        8472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human Myc-PD-L1 in mamalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8472)
  AUTHORS   Li CW, Lim SO, Xia W, Lee HH, Chan LC, Kuo CW, Khoo KH, Chang SS, 
            Cha JH, Kim T, Hsu JL, Wu Y, Hsu JM, Yamaguchi H, Ding Q, Wang Y, 
            Yao J, Lee CC, Wu HJ, Sahin AA, Allison JP, Yu D, Hortobagyi GN, 
            Hung MC
  TITLE     Glycosylation and stabilization of programmed death ligand-1 
            suppresses T-cell activity.
  JOURNAL   Nat Commun. 2016 Aug 30;7:12632. doi: 10.1038/ncomms12632.
  PUBMED    27572267
REFERENCE   2  (bases 1 to 8472)
  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: "Nat 
            Commun."; date: "2016-08-30"; volume: "7:12632. doi"; pages: " 
            10.1038/ncomms12632"
COMMENT     Sequence Label: pCDH-Neo/Myc PD-L1
FEATURES             Location/Qualifiers
     source          1..8472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1228..1257
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        1301..1512
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1525..1793
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1820..2614
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1874..1893)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2484..2503
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2621..3209
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2674..2694)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3092..3103)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3283..3516
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3588..3709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3625..3644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3679..3698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3749..3884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3917..3933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3917..3933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3930..3952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3941..3957
                     /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)."
     promoter        complement(join(3965..3971,3972..3989,3990..3995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4010..4031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4148..4165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4319..4907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5078..5869,5870..5938))
                     /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     5701..5720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5939..6043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6111..6129
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6167..6189)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6289..6308
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6502..6524
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6516..6533
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6517..6533
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6548..6774
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6775..6955
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7002..7127
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7620..7853
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8038..8082
                     /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    8349..8465
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctrgyattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
       61 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      121 gcggtttgac tcaccctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
      181 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
      241 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
      301 tctgcgccgt tacagatcca agctgtgacc ggcgcctact ctagagctag cgaattcatg
      361 aggatatttg ctgtctttat attcatgacc tactggcatt tgctgaacgc atttactgtc
      421 acggttccca aggacctata tgtggtagag tatggtagca atatgacaat tgaatgcaaa
      481 ttcccagtag aaaaacaatt agacctggct gcactaattg tctattggga aatggaggat
      541 aagaacatta ttcaatttgt gcatggagag gaagacctga aggttcagca tagtagctac
      601 agacagaggg cccggctgtt gaaggaccag ctctccctgg gaaatgctgc acttcagatc
      661 acagatgtga aattgcagga tgcaggggtg taccgctgca tgatcagcta tggtggtgcc
      721 gactacaagc gaattactgt gaaagtcaat gccccataca acaaaatcaa ccaaagaatt
      781 ttggttgtgg atccagtcac ctctgaacat gaactgacat gtcaggctga gggctacccc
      841 aaggccgaag tcatctggac aagcagtgac catcaagtcc tgagtggtaa gaccaccacc
      901 accaattcca agagagagga gaagcttttc aatgtgacca gcacactgag aatcaacaca
      961 acaactaatg agattttcta ctgcactttt aggagattag atcctgagga aaaccataca
     1021 gctgaattgg tcatcccaga actacctctg gcacatcctc caaatgaaag gactcacttg
     1081 gtaattctgg gagccatctt attatgcctt ggtgtagcac tgacattcat cttccgttta
     1141 agaaaaggga gaatgatgga tgtgaaaaaa tgtggcatcc aagatacaaa ctcaaagaag
     1201 caaagtgata cacatttgga ggagacggaa caaaaactca tctcagaaga ggatctgtaa
     1261 gcggccgcgc ccctctcccy cccccccccc kmccgtyast gggcagagcg cacatcgccc
     1321 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     1381 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     1441 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     1501 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     1561 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     1621 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     1681 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     1741 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     1801 tgaccggcgc ctccyckara tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1861 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     1921 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     1981 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccgcgacggg
     2041 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2101 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2161 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2221 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2281 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2341 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     2401 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2461 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     2521 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     2581 cgccttctat cgccttcttg acgagttctt ctgagtcgac aatcaacctc tggattacaa
     2641 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2701 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2761 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2821 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2881 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2941 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3001 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     3061 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3121 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3181 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     3241 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3301 cccaacgaag ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     3361 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3421 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3481 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     3541 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     3601 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3661 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     3721 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3781 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3841 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     3901 ccaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3961 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4021 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6061 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6121 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6181 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6241 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6301 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6361 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6421 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6481 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     6541 tgacgcgtgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6601 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6661 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6721 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6781 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6841 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6901 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     6961 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     7021 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     7081 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     7141 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     7201 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     7261 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     7321 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7381 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7441 aaagtaagac caccgcacag caagcggcca ctgatcttca gacctggagg aggagatatg
     7501 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7561 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7621 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7681 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7741 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7801 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7861 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     7921 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     7981 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     8041 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     8101 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     8161 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     8221 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     8281 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     8341 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     8401 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaattcaaa
     8461 attttatcga ta
//
