LOCUS       sequence-471840-        6201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HiBiT-HSD17B13 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6201)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6201)
  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            "Unpublished"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: HiBiT-HSD17B13 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6201
                     /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(2147..2165)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2202..2323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2267..2286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2568..2925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2568..2759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2591..2611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2776..2911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2976..3770
                     /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(3030..3049)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3640..3659
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3834..3882
                     /note="synthetic polyadenylation signal"
     promoter        4038..4142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4143..4211,4212..5003)
                     /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(4361..4380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5161..5749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5241..5260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5865..6148
                     /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 ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgaaca
     1141 ttattcttga aatccttctc cttctgatca ctatcatata tagttacctt gaaagtttgg
     1201 taaaattttt cattcctcaa cggcggaagt ctgtggccgg tgagatagtc ctgattactg
     1261 gcgcaggaca tggaatcggc cgccagacga cgtacgagtt tgctaagcga caatccatac
     1321 tggtactctg ggatattaac aagcggggtg tagaagagac agcagccgag tgtcgcaagc
     1381 tgggcgtgac tgcacacgca tatgtcgtgg actgcagtaa tcgggaggag atttaccgat
     1441 cccttaacca ggtaaagaag gaggtaggag atgtaacaat agttgtaaat aatgccggaa
     1501 cggtctatcc tgcggatctt ctcagtacaa aggatgaaga aatcacaaaa acttttgaag
     1561 tgaatatact ggggcacttc tggataacaa aggccctcct tcccagtatg atggaacgga
     1621 accatggcca catcgtaacc gttgcgtccg tgtgcggaca tgagggcata ccatatttga
     1681 ttccctattg cagcagtaag tttgcggccg tcggttttca caggggattg acaagtgaac
     1741 tgcaagcgtt gggaaagacg ggtattaaaa cctcctgcct ttgtcccgtg tttgtcaaca
     1801 caggctttac gaagaacccc agcacacgat tgtggccggt gctcgagacc gatgaggtag
     1861 tgcggagcct tatagatggt atcctcacca acaaaaaaat gatctttgtt ccctcatata
     1921 tcaatatttt tctgcgattg caaaaatttc tgcctgagcg ggctagtgcg atactgaacc
     1981 ggatgcaaaa catacaattc gaggcggtcg tcggtcacaa gataaagatg aaagtttaaa
     2041 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2101 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2161 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2221 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2281 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2341 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2401 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     2461 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     2521 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     2581 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2641 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2761 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2821 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2881 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct
     2941 gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg
     3001 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3061 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3121 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3181 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3241 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3301 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3361 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3421 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3481 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3541 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3601 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3661 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3721 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3781 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa
     3841 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     3901 tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     3961 gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca
     4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4321 ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat
     4381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta
     4561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4981 cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc
     5041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5581 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     5761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5821 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     5881 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     5941 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     6001 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     6061 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     6121 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     6181 cggcgtagag gatcgagatc t
//
