LOCUS       sequence-472395-        6170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CHRM1-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 6170)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6170)
  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            "Unpublished"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: CHRM1-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(822..840)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(823..840)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        960..1263
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1418..1438
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1471..1518
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1550..2344
                     /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(1604..1623)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2214..2233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(2502..3090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2582..2601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3216..3264
                     /note="synthetic polyadenylation signal"
     CDS             complement(3288..4325)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3301..3309
                     /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(4356..4713)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4370..4505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4522..4713
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4670..4690
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4785..4866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4810..4829
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcgacagcag cggcctggag gagtctctgg tgagccctca gcccctggct gagacctctc
       61 tgagctgctg ctgctgccac cccctttgcc tggcgtctcg gagccctgaa gggctgccag
      121 ctcccgtgct cggttctctg tctcccggta gatgcgccag tagagcgtgc acatgactgt
      181 gacagggagg tagaaggcag ccatggctgt gccaaaggtg atgatgggct gggagaggaa
      241 ctggatgtag cactgcccag ctagcactgt ccgctcccct accaggtact gccagaagag
      301 gatggctggg gcccagagca caaaggaaac cagccaggcc aggccgatca tcagagctgc
      361 ccggcggggt gtgcgcttgg cacggtagct caggggccga gtcacggaga agtagcggtc
      421 aaagctgatg agcagcagat tcatgacgga ggcattgctg gccacatagt ccagggccag
      481 ccagaggtca caagccagcg tgcccagagc ccagtggccc atgagcaggt acgtggtata
      541 gaggttcatg gagaaggtac cgatgatgag gtcagcacag gccaggctca gcaggaagta
      601 gttattgact gtcttgagct ccgtgttgac cttgaaagag atgagtacca gcaggttgcc
      661 tgtcactgtg gctagcgaca ggaggcccgt ggtgatccca atgaaggcca cttgccaggg
      721 accctttcct ggtgccagga cggtgatgtt ggggctgaca gcaggtgggg ctgaagtgtt
      781 catggcgatc gctagctgca aaaagaacaa gtagcttgta ttctatagtg tcacctaaat
      841 gggcgatctg acggttcact aaaccagctc tgcttatata ggcctccctg gcgcgccact
      901 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      961 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1021 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     1081 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1141 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1201 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1261 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
     1321 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     1381 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta
     1441 cggtgggagg tcaaatcgaa ttctaatact gttgacaatt aatcatcggc atagtatatc
     1501 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     1561 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     1621 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     1681 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     1741 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     1801 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     1861 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     1921 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     1981 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2041 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     2101 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     2161 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     2221 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     2281 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     2341 ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     2401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2941 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3061 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     3121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3181 atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg
     3241 taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagtcta aactcgacct
     3301 acctccttag cgcgcggtcg tgtactgggc cggcggttgc cgctgtcggc cagcacctcg
     3361 acgcagccgt cggtccatac ggctgcgctc cggcgagcga tttgtgtgcg accgacggtg
     3421 ccggccccgc tgcggacgat ggcatcgcag cggccttgag cccaggcagc atcgtcgaag
     3481 ttgccgtcca cgaggctctg gtagagctga tccaggccga tgcgcagcat gtaggctcgc
     3541 agacgagggc tgccggccag ctcgggatgc cggcgctcga agtagcgagt ctgctgctcc
     3601 atgcaagcca gccagggccg ccagaagaag atgttggcca cctcgtactg actgtccccg
     3661 aacatagctt cggaccagtc gattacggcg gtgatgcggc cgttgtcggt caggacgttg
     3721 ttgctgccga agtcggcatg gacgaggtgg cgcacctcgg gacagtcttc ggcccacagc
     3781 atcagttcgt ccagggcttg agctacgctg gcggacacgg tgtcgtccat cacggtctgc
     3841 cagtggtaga catggggatc agcaatggcg caaatgaaat cccgccaggt tgtgtactgg
     3901 ccgatgcctt ggggcccgaa tgggccgaag ccgctggttt gactgaggtc ggcggcggca
     3961 atagcatcca tagcttcggc gacaggctgt aacacagcag gcagctctgt ttcggggagg
     4021 tcttggagag taacgccttg ggcgcgtcta ctgatgcagt atgtcaggct ctcgctgaac
     4081 tcgccgatgt ccaacacttc ggggatgggt agtgcagcgc tggcgaagtg gcggtacacg
     4141 tagcggtctt tgtagaagcc atcagcgcag ctattcaccc gcagtacata gccgcgtccg
     4201 ccgacatcga agctgaaggc tcggctctct tcgccctccg acaactgcat caggtcgctc
     4261 acactgtcga acttctcgat gagaaatttt tcaacgctgg tagcggtgag ttcgggcttc
     4321 ttcatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag gcctccaaaa
     4381 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     4441 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     4501 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     4561 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     4621 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     4681 cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca
     4741 gaggttattt caggccatgg tgctgcgcaa gacgcgtgac gtcggcagtg aaaaaaatgc
     4801 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4861 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     4921 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tccgcggccg
     4981 caatacgtcg acgttatcag ctgacttcgt acgagagcct aggtgtttct agagtcgcgg
     5041 ccttagctgt tgatggttac tcggaacagc atggagccgt cgggggtgat caggcgctcg
     5101 tcgataattt tgttgccgtt ccacagggtc cctgttacag tgatcttttt gccgtcgaac
     5161 acggcgatgc cttcatacgg ccgtccgaaa tagttcagca tgttcggcgt aaccccgtcg
     5221 attaccagtg tgccataggg caggatcacc ttaaagtgat gatcatccac agggtacacc
     5281 accttaaaca cctcttcgat ctgggccatt tggtcggcgc tcagaccttc atacgggatg
     5341 atgacatgga tgtcgatctt cagggcattt tcaccgctcc ggacaatcct ttggatcgga
     5401 gttacggaca cggcgagatt ctgcagcaaa ctggacacac ctccctgttc aaggacttgg
     5461 tccaggttgt aggcggctgt ctgttcccag tccccaacga aatcttcgag tgtgaagaca
     5521 ccgctcgagc ctccacctcc gctcccgcca ccacctgaac tgccttgaga aacgcattgg
     5581 cgggagggag tgcggtgcac ggagccaggg cgcttgggga tcttgcgcca gcgtctcttg
     5641 tcccagcggc aaagcagcag caggcgaaag gtgtcccgga aggctttgtt gcagagtgcg
     5701 tagcacatgg ggttgatggt gctgttgacg tagcacagcc agtagcccag ctcccacagg
     5761 gtctcgggaa cacagtcctt gcagaaggtg gacaccagca ccatgatgtt gtacggtgtc
     5821 caggtgagga tgaaggccag gaggatggca ctcagggtcc gagccgcctt cttctccttg
     5881 accagcgaga aggtcttccg cttggccagc tgctcctttc cacggggctt ctggcccttg
     5941 ccagctcgat cacgcccttt cttagtcggc ctcttgactg tatttgggga gctccgtggg
     6001 ggctgcttgg tgggggcctg tgcctcgggg tccaccattg gcatcttgat caccacttcg
     6061 gagccaggct cctctccctc tgaggatgtg agggactcca tggagccttc gtcctcttcc
     6121 tcttcttcct tccagctgta ggcctgcagc agcctggggg cccggcagca
//
