LOCUS       sequence-479818-        6707 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express PINK1-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 6707)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PINK1-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..6707
                     /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(2587..2605)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2588..2605)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2725..3028
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3183..3203
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3236..3283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3315..4109
                     /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(3369..3388)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3979..3998
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4326..4871
                     /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     4772..4791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4988..5005
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5155..5203
                     /note="synthetic polyadenylation signal"
     CDS             complement(5227..6264)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5240..5248
                     /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(6295..6652)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6309..6444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6461..6652
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6609..6629
                     /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 gaaaccaggg cggcgcgcca gctgcacagc agcagggcgg
      841 cctggcacag ggtctcgcac tccagattgg ccagaaacag catcttcatc ttggtctcca
      901 cgcagcactt ctctgtcagc ctgtttgcca gcagggtggc ggcggactgc tgcagcagcc
      961 atcccaccat cttatccagc ttcagattct tcagggccag gatgtgctcg ccccacaggc
     1021 tcaggtgcag cacgtttgct gccactctgg cggatggcct cttgcttgcc tctctctgca
     1081 gcagggcgcg caccagctgc ctcacgtcag gtggcacgga ctctggcagg gctggcagct
     1141 gggcctcctg ataagaccgg gactccaggt gtgccttgcc ctgtccgtag aagggattca
     1201 ccaggccgaa gatctcgtag gcgatggctc ccactgccca tgcatctgcc ttagaatagt
     1261 cgatcacggc tctggggcct ggccttgctg tggacacctc tggtgccatc agacatccgt
     1321 ttcctcctct atccacgtac cagctagaaa atggcagctg caggccgatg ctctcatcgg
     1381 ccagacagca gccgaagtcg gcgatgacca gccatgggca gccatcaggg tccagctcca
     1441 ccaggatatt atcagacttc aggtcccggt gtgcgattcc ctgctgcacc aggtggtcca
     1501 ctccctccag cagctgcagc agcatcatgg cggccagtct aggagagggg gtattcacgc
     1561 acaggtactg ccgcagtgtg caagggtagt tcttcatgac caggaacagg gtccgtccgt
     1621 gtcccagtcc ctctgggtgc agcctggagg gcagcacatc tgggtagtcc accagggcgc
     1681 caggcagcag tggcacggag cttgtgaagg ctctcagcac ccggatgata tttgggtgag
     1741 gtgccagctg cttaggtccc ctcttggact tcctataggt cacggcgccg tactcgcctg
     1801 ccagggccac cctggatgct ggcaccagct cctggctcat tgtattcagg atggcctcag
     1861 aggagctgcc ggcgctgatg ttccacatca tcttgatggc cagtgggaag gcaggggcgc
     1921 cgggggctct ctcctggccc tcgccggggg cgctggtgcc aggtcccctt cctggcagca
     1981 ggcctgtaga cttggtcacc tccaggttct gtggcagtgt aggcatggtg gcctcataca
     2041 cggcggcaga acagcccttg ccgatggact ggccgatcag gtactcctcc aggcgaaagc
     2101 cctgcagtct ccgggtatcc agtgggtctg gtccaggctt gctcttctgt gtgaagatgg
     2161 cctggatctc ctggcaggcg ctcacggccc tcctagactc tgcctgcttc tcctcgatca
     2221 ggcccaggcc caggccaaag gccaggaaca cggctctgcc acatggtcct gcgcatcccc
     2281 atgcccgcac cacaaactgc ctctgcagcc ttgctgccag tcctgccaca gactgcctaa
     2341 agaagcgcag cctgtttggc agtcccagtc ccactctccg tggctcggcg ccagggccgg
     2401 cggcccagcc aggtctctcg ccccgcacgc agcctgctgc tggtcctggg cggcccagtc
     2461 cgtaggccct gccaggcttg cctgtgaatc tcagcagcag ggcccgtccc agctgcagtc
     2521 cccttcccag ggcctgcctc acggccatgg cgatcgctag ctgcaaaaag aacaagtagc
     2581 ttgtattcta tagtgtcacc taaatgggcg atctgacggt tcactaaacc agctctgctt
     2641 atataggcct ccctggcgcg ccactagtta ttaatagtaa tcaattacgg ggtcattagt
     2701 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2761 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2821 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2881 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     2941 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     3001 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     3061 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3121 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     3181 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtcaaa tcgaattcta atactgttga
     3241 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3301 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3361 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3421 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3481 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3541 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3601 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3661 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3721 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3781 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3841 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3901 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3961 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4021 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4081 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     4141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4201 aaagaacatg tcaaacatga gaattacaac ttatatcgta tggggctgac ttcaggtgct
     4261 acatttgaag agataaattg cactgaaatc tagaaatatt ttatctgatt aataagatga
     4321 tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc
     4381 ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     4441 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     4501 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     4561 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     4621 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     4681 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     4741 gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg
     4801 tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga
     4861 gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc
     4921 atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac
     4981 cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg
     5041 acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca
     5101 gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggaa tcgtcacaca
     5161 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     5221 tagagtctaa actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc
     5281 gctgtcggcc agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat
     5341 ttgtgtgcga ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc
     5401 ccaggcagca tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat
     5461 gcgcagcatg taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa
     5521 gtagcgagtc tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac
     5581 ctcgtactga ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc
     5641 gttgtcggtc aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg
     5701 acagtcttcg gcccacagca tcagttcgtc cagggcttga gctacgctgg cggacacggt
     5761 gtcgtccatc acggtctgcc agtggtagac atggggatca gcaatggcgc aaatgaaatc
     5821 ccgccaggtt gtgtactggc cgatgccttg gggcccgaat gggccgaagc cgctggtttg
     5881 actgaggtcg gcggcggcaa tagcatccat agcttcggcg acaggctgta acacagcagg
     5941 cagctctgtt tcggggaggt cttggagagt aacgccttgg gcgcgtctac tgatgcagta
     6001 tgtcaggctc tcgctgaact cgccgatgtc caacacttcg gggatgggta gtgcagcgct
     6061 ggcgaagtgg cggtacacgt agcggtcttt gtagaagcca tcagcgcagc tattcacccg
     6121 cagtacatag ccgcgtccgc cgacatcgaa gctgaaggct cggctctctt cgccctccga
     6181 caactgcatc aggtcgctca cactgtcgaa cttctcgatg agaaattttt caacgctggt
     6241 agcggtgagt tcgggcttct tcatggtggc gctagtgtca gaagaatcga gctttttgca
     6301 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     6361 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     6421 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     6481 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     6541 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     6601 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc
     6661 taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgc
//
