LOCUS       sequence-471888-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CD3E-LgBiT 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 5411)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5411)
  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            "Unpublished"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: CD3E-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1465..1483)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1466..1483)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1603..1906
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2061..2081
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2114..2161
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2193..2987
                     /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(2247..2266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2857..2876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3145..3733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3225..3244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3859..3907
                     /note="synthetic polyadenylation signal"
     CDS             complement(3931..4968)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3944..3952
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(4999..5356)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5013..5148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5165..5356
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5313..5333
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaacgatgc gtctctgatt caggccagaa tacaggtccc
      841 gctggccttt ccggatgggc tcatagtctg ggttgggaac aggtggtggc ctctccttgt
      901 tttgtcccct ttgcctgccg ccagcacccg ctcctcgtgt cacaggcttg gccttggcct
      961 ttctattctt gctccagtag taaaccagca gcagcaagcc cccagtgatg cagatgtcca
     1021 ctatgacaat tgtggccacc gacatcacat ccatctccat gcagttctca cacactcttg
     1081 ccctcaggta gagataaaag ttcgcatctt ctggtttgct tcctctgggg tagcagacat
     1141 aataaccact ttgctccaat tctgaaaatt ccttcagtga caggtgatcc tcatcactgc
     1201 ctatgttttt atcatcctca tcaccgccta tgtttttatc attgtgttgc catagtattt
     1261 cagatccagg atactgaggg catgtcaata ttactgtggt tccagagatg gagactttat
     1321 atggtgtctg tgtaatacca cccatttctt cattaccatc ttgcccccaa acgccaactg
     1381 ataagaggca gaggcccaga actctccagt gagtgcccga ctgcatggcg atcgctagct
     1441 gcaaaaagaa caagtagctt gtattctata gtgtcaccta aatgggcgat ctgacggttc
     1501 actaaaccag ctctgcttat ataggcctcc ctggcgcgcc actagttatt aatagtaatc
     1561 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1621 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1681 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1741 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     1801 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     1861 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     1921 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     1981 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2041 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtcaaatc
     2101 gaattctaat actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     2161 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     2221 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2281 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2341 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2401 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2461 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2521 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2581 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2641 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2701 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2761 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2821 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2881 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2941 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     3001 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3721 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3841 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     3901 ttttattgcg gccatcgtga tggctagagt ctaaactcga cctacctcct tagcgcgcgg
     3961 tcgtgtactg ggccggcggt tgccgctgtc ggccagcacc tcgacgcagc cgtcggtcca
     4021 tacggctgcg ctccggcgag cgatttgtgt gcgaccgacg gtgccggccc cgctgcggac
     4081 gatggcatcg cagcggcctt gagcccaggc agcatcgtcg aagttgccgt ccacgaggct
     4141 ctggtagagc tgatccaggc cgatgcgcag catgtaggct cgcagacgag ggctgccggc
     4201 cagctcggga tgccggcgct cgaagtagcg agtctgctgc tccatgcaag ccagccaggg
     4261 ccgccagaag aagatgttgg ccacctcgta ctgactgtcc ccgaacatag cttcggacca
     4321 gtcgattacg gcggtgatgc ggccgttgtc ggtcaggacg ttgttgctgc cgaagtcggc
     4381 atggacgagg tggcgcacct cgggacagtc ttcggcccac agcatcagtt cgtccagggc
     4441 ttgagctacg ctggcggaca cggtgtcgtc catcacggtc tgccagtggt agacatgggg
     4501 atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac tggccgatgc cttggggccc
     4561 gaatgggccg aagccgctgg tttgactgag gtcggcggcg gcaatagcat ccatagcttc
     4621 ggcgacaggc tgtaacacag caggcagctc tgtttcgggg aggtcttgga gagtaacgcc
     4681 ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg aactcgccga tgtccaacac
     4741 ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac acgtagcggt ctttgtagaa
     4801 gccatcagcg cagctattca cccgcagtac atagccgcgt ccgccgacat cgaagctgaa
     4861 ggctcggctc tcttcgccct ccgacaactg catcaggtcg ctcacactgt cgaacttctc
     4921 gatgagaaat ttttcaacgc tggtagcggt gagttcgggc ttcttcatgg tggcgctagt
     4981 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     5041 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5101 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5161 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5221 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5281 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5341 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     5401 tggtgctgcg c
//
