LOCUS       sequence-474958-        5545 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express PD-L1-HaloTag(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 5545)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PD-L1-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5545
                     /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..1976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1992..2876
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(2964..2982)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3019..3140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3030..3049)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3084..3103
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3539..4333
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3593..3612)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4203..4222
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4505..5093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4585..4604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5209..5492
                     /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 ctcgagccaa ccactgagga tctgtacttt cagagcgata
     1981 acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
     2041 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
     2101 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
     2161 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
     2221 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
     2281 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
     2341 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
     2401 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
     2461 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
     2521 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     2581 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     2641 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     2701 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     2761 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     2821 cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt tccggttaat
     2881 agaattctag agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa
     2941 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc
     3001 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3061 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3121 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3181 gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat
     3241 tctaatggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     3301 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc
     3361 agcccttgcg ccctgagtgc ttgcggcagc gtgagcttca aaagaattgc cagctggggc
     3421 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     3481 gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgacggt cgtttcgcat
     3541 gcttgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3601 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3661 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3721 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3781 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3841 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3901 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3961 cgagcgagca cgcactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4021 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgta tgccggatgg
     4081 tgaggatctc gtcgtgactc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4141 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4201 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4261 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4321 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaaccgg
     4381 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4441 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4501 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4561 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4621 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4681 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4741 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4801 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4861 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4921 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4981 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5041 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     5101 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5161 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag
     5221 gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag
     5281 tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta
     5341 aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc
     5401 tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa
     5461 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     5521 cgtccggcgt agaggatcga gatct
//
