LOCUS       sequence-478483-        5358 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express CCR2-HiBiT 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 5358)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5358)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CCR2-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5358
                     /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"
     primer_bind     complement(2301..2319)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2356..2477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2367..2386)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2421..2440
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2722..3079
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2722..2913
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2745..2765)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2930..3065
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3094..3141
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3173..3967
                     /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(3227..3246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3837..3856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4031..4079
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4318..4906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4398..4417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5022..5305
                     /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 cgccatgctg tccacatctc
     1081 gttctcggtt tatcagaaat accaacgaga gcggtgaaga agtcaccacc ttttttgatt
     1141 atgattacgg tgctccctgt cataaatttg acgtgaagca aattggggcc caactcctgc
     1201 ctccgctcta ctcgctggtg ttcatctttg gttttgtggg caacatgctg gtcgtcctca
     1261 tcttaataaa ctgcaaaaag ctgaagtgct tgactgacat ttacctgctc aacctggcca
     1321 tctctgatct gctttttctt attactctcc cattgtgggc tcactctgct gcaaatgagt
     1381 gggtctttgg gaatgcaatg tgcaaattat tcacagggct gtatcacatc ggttattttg
     1441 gcggaatctt cttcatcatc ctcctgacaa tcgatagata cctggctatt gtccatgctg
     1501 tgtttgcttt aaaagccagg acggtcacct ttggggtggt gacaagtgtg atcacctggt
     1561 tggtggctgt gtttgcttct gtcccaggaa tcatctttac taaatgccag aaagaagatt
     1621 ctgtttatgt ctgtggccct tattttccac gaggatggaa taatttccac acaataatga
     1681 ggaacatttt ggggctggtc ctgccgctgc tcatcatggt catctgctac tcgggaatcc
     1741 tgaaaaccct gcttcggtgt cgaaacgaga agaagaggca tagggcagtg agagtcatct
     1801 tcaccatcat gattgtttac tttctcttct ggactcccta taatattgtc attctcctga
     1861 acaccttcca ggaattcttc ggcctgagta actgtgaaag caccagtcaa ctggaccaag
     1921 ccacgcaggt gacagagact cttgggatga ctcactgctg catcaatccc atcatctatg
     1981 ccttcgttgg ggagaagttc agaaggtatc tctcggtgtt cttccgaaag cacatcacca
     2041 agcgcttctg caaacaatgt ccagttttct acagggagac agtggatgga gtgacttcaa
     2101 caaacacgcc ttccactggg gagcaggaag tctcggctgg tttagtttct caaggcagtt
     2161 caggtggctc gagcggtgtg agcggctggc ggctgttcaa gaagattagc taaggccgcg
     2221 actctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2281 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2341 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2401 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2461 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2521 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2581 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2641 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2701 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2761 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2821 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2881 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2941 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3001 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3061 aggcctaggc ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat
     3121 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     3181 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3241 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3301 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3361 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3421 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3481 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3541 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3601 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3661 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3721 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3781 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3841 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3901 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3961 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4021 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     4081 atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     4141 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacc cggtatcagc
     4201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4741 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat
     4921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4981 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
     5041 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
     5101 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
     5161 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
     5221 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
     5281 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg
     5341 cgtagaggat cgagatct
//
