LOCUS       sequence-473368-        6128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HiBiT-TDP1 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6128)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6128)
  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            "Unpublished"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-TDP1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6128
                     /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(3071..3089)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3126..3247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3137..3156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3191..3210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3492..3849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3492..3683
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3515..3535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3700..3835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3864..3911
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3943..4737
                     /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(3997..4016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4607..4626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4801..4849
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5088..5676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5168..5187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5792..6075
                     /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 gccatgtctc
     1141 aggaaggcga ttatgggagg tggaccatat ctagtagtga tgaaagtgag gaagaaaagc
     1201 caaaaccaga caagccatct acctcttctc ttctctgtgc caggcaagga gcagcaaatg
     1261 agcccaggta cacctgttcc gaggcccaga aagctgcaca caagaggaaa atatcacctg
     1321 tgaaattcag caatacagat tcagttttac ctcccaaaag gcagaaaagc ggttcccagg
     1381 aggacctcgg ctggtgtctg tccagcagtg atgatgagct gcaaccagaa atgccgcaga
     1441 agcaggctga gaaagtggtg atcaaaaagg agaaagacat ctctgctccc aatgacggca
     1501 ctgcccaaag aactgaaaat catggcgctc ccgcctgcca caggctcaaa gaggaggaag
     1561 acgagtatga gacatcaggg gagggccagg acatttggga catgctggat aaagggaacc
     1621 ccttccagtt ttacctcact agagtctctg gagttaagcc aaagtataac tctggagccc
     1681 tccacatcaa ggatatttta tctcctttat ttgggacgct tgtttcttca gctcagttta
     1741 actactgctt tgacgtggac tggctcgtaa aacagtatcc accagagttc aggaagaagc
     1801 caatcctgct tgtgcatggt gataagcgag aggctaaggc tcacctccat gcccaggcca
     1861 agccttacga gaacatctct ctctgccagg caaagttgga tattgcgttt ggaacacacc
     1921 acacgaaaat gatgctgctg ctctatgaag aaggcctccg ggttgtcata cacacctcca
     1981 acctcatcca tgctgactgg caccagaaaa ctcaaggaat atggttgagc cccttatacc
     2041 cacgaattgc tgatggaacc cacaaatctg gagagtcgcc aacacatttt aaagctgatc
     2101 tcatcagtta cttgatggct tataatgccc cttctctcaa ggagtggata gatgtcattc
     2161 acaagcacga tctctctgaa acaaatgttt atcttattgg ttcaacccca ggacgctttc
     2221 aaggaagtca aaaagataat tggggacatt ttagacttaa gaagcttctg aaagaccatg
     2281 cctcatccat gcctaacgca gagtcctggc ctgtcgtagg tcagttttca agcgttggct
     2341 ccttgggagc cgatgaatca aagtggttat gttctgagtt taaagagagc atgctgacac
     2401 tggggaagga aagcaagact ccaggaaaaa gctctgttcc tctttacttg atctatcctt
     2461 ctgtggaaaa tgtgcggacc agtttagaag gatatcctgc tgggggctct cttccctata
     2521 gcatccagac agctgaaaaa cagaattggc tgcattccta ttttcacaaa tggtcagctg
     2581 agacttctgg ccgcagcaat gccatgccac atattaagac atatatgagg ccttctccag
     2641 acttcagtaa aattgcttgg ttccttgtca caagcgcaaa tctgtccaag gctgcctggg
     2701 gagcattgga gaagaatggc acccagctga tgatccgctc ctacgagctc ggggtccttt
     2761 tcctcccttc agcatttggt ctagacagtt tcaaagtgaa acagaagttc ttcgctggca
     2821 gccaggagcc aatggccacc tttcctgtgc catatgattt gcctccagaa ctgtatggaa
     2881 gtaaagatcg gccatggata tggaacattc cttatgtcaa agcaccggat acgcatggga
     2941 acatgtgggt gccctccgtt taaacgaatt cgggctcggt acccggggat cctctagagt
     3001 cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     3061 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     3121 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3181 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3241 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3301 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     3361 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3421 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3481 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3541 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3601 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3661 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3721 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3781 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3841 ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag
     3901 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3961 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4021 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4081 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4141 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4201 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4261 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4321 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4381 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4441 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4501 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4561 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4621 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4681 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4741 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4801 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4861 ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     4921 atccggggtc agcaccgttt ctgcggactg gctttctacc cggtatcagc tcactcaaag
     4981 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5521 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5641 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     5701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5761 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg
     5821 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta
     5881 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga
     5941 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc
     6001 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     6061 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat
     6121 cgagatct
//
