LOCUS       sequence-471841-        6175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HSD17B13-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6175)
  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            "Unpublished"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: HSD17B13-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6175
                     /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"
     primer_bind     complement(2121..2139)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2176..2297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2187..2206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2241..2260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2542..2899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2542..2733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2565..2585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2750..2885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2950..3744
                     /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(3004..3023)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3614..3633
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3808..3856
                     /note="synthetic polyadenylation signal"
     promoter        4012..4116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4117..4185,4186..4977)
                     /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(4335..4354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5135..5723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5215..5234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5839..6122
                     /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 cgccatgaac attattcttg
     1081 aaatccttct ccttctgatc actatcatat atagttacct tgaaagtttg gtaaaatttt
     1141 tcattcctca acggcggaag tctgtggccg gtgagatagt cctgattact ggcgcaggac
     1201 atggaatcgg ccgccagacg acgtacgagt ttgctaagcg acaatccata ctggtactct
     1261 gggatattaa caagcggggt gtagaagaga cagcagccga gtgtcgcaag ctgggcgtga
     1321 ctgcacacgc atatgtcgtg gactgcagta atcgggagga gatttaccga tcccttaacc
     1381 aggtaaagaa ggaggtagga gatgtaacaa tagttgtaaa taatgccgga acggtctatc
     1441 ctgcggatct tctcagtaca aaggatgaag aaatcacaaa aacttttgaa gtgaatatac
     1501 tggggcactt ctggataaca aaggccctcc ttcccagtat gatggaacgg aaccatggcc
     1561 acatcgtaac cgttgcgtcc gtgtgcggac atgagggcat accatatttg attccctatt
     1621 gcagcagtaa gtttgcggcc gtcggttttc acaggggatt gacaagtgaa ctgcaagcgt
     1681 tgggaaagac gggtattaaa acctcctgcc tttgtcccgt gtttgtcaac acaggcttta
     1741 cgaagaaccc cagcacacga ttgtggccgg tgctcgagac cgatgaggta gtgcggagcc
     1801 ttatagatgg tatcctcacc aacaaaaaaa tgatctttgt tccctcatat atcaatattt
     1861 ttctgcgatt gcaaaaattt ctgcctgagc gggctagtgc gatactgaac cggatgcaaa
     1921 acatacaatt cgaggcggtc gtcggtcaca agataaagat gaaagtttct caaggcagtt
     1981 caggtggctc gagcggtgtg agcggctggc ggctgttcaa gaagattagc taaggccgcg
     2041 actctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2101 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2161 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2221 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2281 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2341 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2401 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2461 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2521 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2581 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2641 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2701 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2761 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2821 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2881 aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct
     2941 agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3001 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3061 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3121 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3181 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3241 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3301 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3361 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3421 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3481 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3541 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3601 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3661 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3721 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3781 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     3841 gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt
     3901 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     3961 ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4021 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4321 actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4501 gataacactg cggccaactt acttctgaca actatcggag gaccgaagga gctaaccgct
     4561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4621 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaattc
     4981 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     5041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5701 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5821 cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga
     5881 tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac
     5941 cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt
     6001 tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta
     6061 gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc
     6121 gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
