LOCUS       sequence-474957-        5613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PD-L1-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5613)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5613)
  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            "Unpublished"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: PD-L1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1956..2468
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2556..2574)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2611..2732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2622..2641)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2676..2695
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2977..3334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2977..3168
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3000..3020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3185..3320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3349..3396
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3428..4222
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3482..3501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4092..4111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4286..4334
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4573..5161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4653..4672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5277..5560
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgagg atatttgctg
     1081 tctttatatt catgacctac tggcatttgc tgaacgcatt tactgtcacg gttcccaagg
     1141 acctatatgt ggtagagtat ggtagcaata tgacaattga atgcaaattc ccagtagaaa
     1201 aacaattaga cctggctgca ctaattgtct attgggaaat ggaggataag aacatcattc
     1261 aatttgtgca tggagaggaa gacctgaagg ttcagcatag tagctacaga cagagggccc
     1321 ggctgttgaa ggaccagctc tccctgggaa atgctgcact tcagatcaca gatgtgaaat
     1381 tgcaggatgc aggggtgtac cgctgcatga tcagctatgg tggtgccgac tacaagcgaa
     1441 ttactgtgaa agtcaatgcc ccatacaaca aaatcaacca aagaattttg gttgtggatc
     1501 cagtcacctc tgaacatgaa ctgacatgtc aggctgaggg ctaccccaag gccgaagtca
     1561 tctggacaag cagtgaccat caagtcctga gtggtaagac caccaccacc aattccaaga
     1621 gagaggagaa gcttttcaat gtgaccagca cactgagaat caacacaaca actaatgaga
     1681 ttttctactg cacttttagg agattagatc ctgaggaaaa ccatacagct gaattggtca
     1741 tcccagaact acctctggca catcctccaa atgaaaggac tcacttggta attctgggag
     1801 ccatcttatt atgccttggt gtagcactga cattcatctt ccgtttaaga aaagggagaa
     1861 tgatggatgt gaaaaaatgt ggcatccaag atacaaactc aaagaagcaa agtgatacac
     1921 atttggagga gacggtttct ctcggctcga gcggcgtctt cacactcgaa gatttcgttg
     1981 gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt
     2041 ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg
     2101 gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg
     2161 accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact
     2221 ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg
     2281 actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa
     2341 cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct
     2401 ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc
     2461 tggcgtaagg ccgcgactct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     2521 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2581 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2641 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2701 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2761 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     2821 gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     2881 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     2941 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3241 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3301 aggaggcttt tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa
     3361 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag
     3421 gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3481 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3541 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3601 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3661 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3721 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3781 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3841 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3901 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3961 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4021 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4081 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4141 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4201 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4261 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4321 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     4381 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     4441 ctacccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4981 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     5161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5221 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     5281 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     5341 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     5401 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     5461 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     5521 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     5581 ccacgatgcg tccggcgtag aggatcgaga tct
//
