LOCUS       sequence-479828-        7001 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express LgBiT-Syk 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 7001)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7001)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LgBiT-Syk BiBiT Ready Vector
FEATURES             Location/Qualifiers
     source          1..7001
                     /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"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(2881..2899)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2882..2899)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3019..3322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3477..3497
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3530..3577
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3609..4403
                     /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(3663..3682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4273..4292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4620..5165
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5066..5085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5282..5299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5449..5497
                     /note="synthetic polyadenylation signal"
     CDS             complement(5521..6558)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5534..5542
                     /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(6589..6946)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6603..6738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6755..6946
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6903..6923
                     /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 ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaacgttc accacgtcat
      421 agtagtaatt gcgcagccgc agttccactg ctgcgaatcc gggcctgttt tccacatcgt
      481 atgtccagca cagattcatg agatcgtaca tctctcttgg acaccctgca gggcacccca
      541 tccgctctcc tttctctaac atagcggtga cttcacttcc tttcatccct cgatatggct
      601 tctgcccata ggagaatgct tcccacatca acactccaaa gctccagaca tcgcttttgc
      661 tggagaactt gtagtagttg atgcattccg gagcgtacca cttgacaggc cactttccat
      721 gggtctgggc cttgtagtag ttttcatcag cacgcagtgc tttggaaagt ccgaaatcac
      781 tgatcttggc gtaatgttgg gtaactagca acacatttct tgcagccaga tctctgtgca
      841 caaaattgct ctcctccaag tacttcatgc ccatggaaac ctgatgaacc agttctatga
      901 tgttcttatc cttgacatgt ctgttctgct gcaaatactt attgagggga ccaagttctg
      961 ccatctccat aaccagcatc caggactcgg cctcgcatat cccgatcatc cgcacgatgt
     1021 acgggttgtc cagctgctgc atgacatttg cttctgctaa taactcatct ttaagagcgg
     1081 ggtcattggc ctcgtttttc agtattttca cagccacggt tttcacaact tttttcattt
     1141 ggtagtagcc ctttttcaca gttccaaaat taccagagcc cagttctttg tcttccagcg
     1201 tcagcagctt tcggtccagg taaacctcct tgggcctgat ctcctcgggg tccgcgtagg
     1261 ggctctcgta cacctctgtg tccatgggta gggcttctct ctgggggcct ttgtctgcag
     1321 cccagggtgc aagttctggc tcatacggat tgaatgacac agtactctct tgccggttcc
     1381 cttgggcagg ggaggacttt ctgtggccag gctttgggaa ggagtatgat ttgattcttg
     1441 agattattcc acccgctgac caagtcgcag gatgggaacc tggaagttgt ggacggcctc
     1501 caaaattaac atttccctgt gtgccgattt tttgacatgg gacagtaaga actcttaaca
     1561 aaccatctgc tttataagaa taatgctcga ctagctgcca gagcgtgtcg aacttctttc
     1621 cctcggggat ggagagcttc cctgtcttgt ctttgtcgat gcgatagtgc agcaccttcc
     1681 cttcgtgcag caggcacagg gcgtaggagc cgttgttgtc tctggctcgg atcaggaact
     1741 ttccatttgt ctttgatcct atcaggacaa tttgctcaga ttcttcccga gagatttttc
     1801 catggaacca aggcattttt tcatgggctg tggtagcgat cagcttctcc agctgaggct
     1861 tctgactgat gatggcctgc tccagagcct gaccctgcag gttccatgtc tgcttcacat
     1921 attccctgat gaggttttcc ttcaaatcct caaagggccc agtcttgggc tgcacccctt
     1981 ggggccggtt gaagggcttc ttgaggaggc agaccaggcc atcagactcc tgggagtggt
     2041 agtggcagag gtcggcgggg ctggcatggg tcctgccacc ggcgatggcg taggtgccat
     2101 tcagctcccg ctcgatggtg tagtggtgtg ccttcctccc gtgggccacg gacagggcga
     2161 agccacccag gtagttgcgg ctctggcgca gcaaataaag cccatcactc atgcccccct
     2221 ggaccaggta atcttctgcc tcctcccggg tgatgttgcc gaaaaagaag ggcaggtggt
     2281 tggcgctgtc agccatgccg ctgctggcca tggcgatcgc accgctcgag cctccacctc
     2341 cgctcccgcc accaccggag ctcccactgt tgatggttac tcggaacagc atggagccgt
     2401 cgggggtgat caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag
     2461 tgatcttttt gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagttcagca
     2521 tgttcggcgt aaccccgtcg attaccagtg tgccataggg caggatcacc ttaaagtgat
     2581 gatcatccac agggtacacc accttaaaca cctcttcgat ctgggccatt tggtcggcgc
     2641 tcagaccttc atacgggatg atgacatgga tgtcgatctt cagggcattt tcaccgctcc
     2701 ggacaatcct ttggatcgga gttacggaca cggcgagatt ctgcagcaaa ctggacacac
     2761 ctccctgttc aaggacttgg tccaggttgt aggcggctgt ctgttcccag tccccaacga
     2821 aatcttcgag tgtgaagacc atggtgagcg ctagctgcaa aaagaacaag tagcttgtat
     2881 tctatagtgt cacctaaatg ggcgatctga cggttcacta aaccagctct gcttatatag
     2941 gcctccctgg cgcgccacta gttattaata gtaatcaatt acggggtcat tagttcatag
     3001 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     3061 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     3121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     3181 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     3241 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     3301 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     3361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     3481 aatgggcggt aggcgtgtac ggtgggaggt caaatcgaat tctaatactg ttgacaatta
     3541 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     3601 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3661 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3721 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3781 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3841 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3901 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3961 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4021 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4081 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4141 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4201 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4261 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4321 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4381 gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc
     4441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4501 catgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     4561 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     4621 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     4681 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     4741 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     4801 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     4861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     4921 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     4981 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     5041 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5101 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     5161 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     5221 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     5281 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     5341 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     5401 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggaatcgtca cacaaaaaac
     5461 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt
     5521 ctaaactcga cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc
     5581 ggccagcacc tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt
     5641 gcgaccgacg gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc
     5701 agcatcgtcg aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag
     5761 catgtaggct cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg
     5821 agtctgctgc tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta
     5881 ctgactgtcc ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc
     5941 ggtcaggacg ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc
     6001 ttcggcccac agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc
     6061 catcacggtc tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca
     6121 ggttgtgtac tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag
     6181 gtcggcggcg gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc
     6241 tgtttcgggg aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag
     6301 gctctcgctg aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa
     6361 gtggcggtac acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac
     6421 atagccgcgt ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg
     6481 catcaggtcg ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt
     6541 gagttcgggc ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc
     6601 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     6661 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     6721 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     6781 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     6841 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6901 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     6961 agttcctctt tcagaggtta tttcaggcca tggtgctgcg c
//
