LOCUS       sequence-474757-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding HaloTag(R)-LgBiT for use in 
            HiBiT cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6811)
  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            "Unpublished"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: CMV HaloTag-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..6811
                     /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"
     CDS             1065..1955
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(2574..2592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2629..2750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2640..2659)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2694..2713
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2995..3352
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2995..3186
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3018..3038)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3203..3338
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3383..4420
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4399..4407
                     /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)"
     polyA_signal    4444..4492
                     /note="synthetic polyadenylation signal"
     promoter        4648..4752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4753..4821,4822..5613)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4971..4990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5771..6359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5851..5870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6475..6758
                     /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 cgccatggca gaaatcggta
     1081 ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg
     1141 atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct
     1201 acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc
     1261 tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc
     1321 gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg
     1381 actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta
     1441 ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc
     1501 gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga
     1561 acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga
     1621 tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc
     1681 caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca
     1741 tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc
     1801 tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg
     1861 acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga
     1921 tcgcgcgctg gctgtcgacg ctcgagattt ccggcgtttc tcaaggcagt tcaggtggtg
     1981 gcgggagcgg aggtggaggc tcgagcggtg tcttcacact cgaagatttc gttggggact
     2041 gggaacagac agccgcctac aacctggacc aagtccttga acagggaggt gtgtccagtt
     2101 tgctgcagaa tctcgccgtg tccgtaactc cgatccaaag gattgtccgg agcggtgaaa
     2161 atgccctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc gccgaccaaa
     2221 tggcccagat cgaagaggtg tttaaggtgg tgtaccctgt ggatgatcat cactttaagg
     2281 tgatcctgcc ctatggcaca ctggtaatcg acggggttac gccgaacatg ctgaactatt
     2341 tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact gtaacaggga
     2401 ccctgtggaa cggcaacaaa attatcgacg agcgcctgat cacccccgac ggctccatgc
     2461 tgttccgagt aaccatcaac agctaaggcc gcgactctag agtcgacctg caggcatgca
     2521 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2581 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     2641 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2701 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2761 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     2821 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     2941 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     3001 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3181 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3241 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3301 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga
     3361 ttcttctgac actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa
     3421 atttctcatc gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga
     3481 gagccgagcc ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg
     3541 cgctgatggc ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc
     3601 catccccgaa gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag
     3661 acgcgcccaa ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca
     3721 gcctgtcgcc gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt
     3781 cggcccattc gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc
     3841 cattgctgat ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag
     3901 cgtagctcaa gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca
     3961 cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc
     4021 cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt
     4081 cttctggcgg ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca
     4141 tcccgagctg gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca
     4201 gctctaccag agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg
     4261 cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag
     4321 cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc
     4381 cagtacacga ccgcgcgcta aggaggtagg tcgagtttag actctagcca tcacgatggc
     4441 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4501 gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     4561 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg
     4621 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4741 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4801 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4861 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4921 agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc
     4981 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5041 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5101 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5161 ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5221 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5281 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5341 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5401 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5461 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5521 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5581 gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc
     5641 aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5881 ccctcgtgcg ctctcctgtt ccgacgctgc cgcttaccgg atacctgtcc gcctttctcc
     5941 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6181 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6301 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag
     6361 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa
     6481 atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag
     6541 cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata
     6601 cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta
     6661 cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta
     6721 tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc
     6781 acgatgcgtc cggcgtagag gatcgagatc t
//
