LOCUS       sequence-478465-        5138 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LDHA-HiBiT 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 5138)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LDHA-HiBiT Vector
FEATURES             Location/Qualifiers
     source          1..5138
                     /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             1235..1246
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2217..2235)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2272..2393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2283..2302)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2337..2356
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2638..2995
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2638..2829
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2661..2681)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2846..2981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3010..3057
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3089..3883
                     /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(3143..3162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3753..3772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3947..3995
                     /note="synthetic polyadenylation signal"
     rep_origin      4152..4697
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4814..4831
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 actctaaagg
     1081 atcagctgat ttataatctt ctaaaggaag aacagacccc ccagaataag attacagttg
     1141 ttggggttgg tgctgttggc atggcctgtg ccatcagtat cttaatgaag gacttggcag
     1201 atgaacttgc tcttgttgat gtcatcgaag acaaattgaa gggagagatg atggatctcc
     1261 aacatggcag ccttttcctt agaacaccaa agattgtctc tggcaaagac tataatgtaa
     1321 ctgcaaactc caagctggtc attatcacgg ctggggcacg tcagcaagag ggagaaagcc
     1381 gtcttaattt ggtccagcgt aacgtgaaca tctttaaatt catcattcct aatgttgtaa
     1441 aatacagccc gaactgcaag ttgcttattg tttcaaatcc agtggatatc ttgacctacg
     1501 tggcttggaa gataagtggt tttcccaaaa accgtgttat tggaagtggt tgcaatctgg
     1561 attcagcccg attccgttac ctgatggggg aaaggctggg agttcaccca ttaagctgtc
     1621 atgggtgggt ccttggggaa catggagatt ccagtgtgcc tgtatggagt ggaatgaatg
     1681 ttgctggtgt ctctctgaag actctgcacc cagatttagg gactgataaa gataaggaac
     1741 agtggaaaga ggttcacaag caggtggttg agagtgctta tgaggtgatc aaactcaaag
     1801 gctacacatc ctgggctatt ggactctctg tagcagattt ggcagagagt ataatgaaga
     1861 atcttaggcg ggtgcaccca gtttccacca tgattaaggg tctttacgga ataaaggatg
     1921 atgtcttcct tagtgttcct tgcattttgg gacagaatgg aatctcagac cttgtgaagg
     1981 tgactctgac ttctgaggaa gaggcccgtt tgaagaagag tgcagataca ctttggggga
     2041 tccaaaagga gctgcaattt gtttctcaag gcagttcagg tggctcgagc ggtgtgagcg
     2101 gctggcggct gttcaagaag attagctaag gccgcgactc tagagtcgac ctgcaggcat
     2161 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2221 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     2281 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2341 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2401 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     2461 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     2521 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     2581 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     2641 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2701 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2821 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2881 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2941 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3001 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3061 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     3121 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3181 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3241 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3301 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3361 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3421 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3481 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3541 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3601 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3661 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3721 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3781 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3841 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3901 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     3961 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc
     4021 atgcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca
     4081 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     4141 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     4201 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     4261 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     4321 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     4381 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     4441 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     4501 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     4561 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     4681 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     4741 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     4801 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     4861 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     4921 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc
     4981 ttttgccgtt acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa
     5041 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     5101 tccggccacg atgcgtccgg cgtagaggat cgagatct
//
