LOCUS       sequence-479508-        5441 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Express HiBiT-GALR1 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 5441)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-GALR1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5441
                     /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(2384..2402)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2439..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2450..2469)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2504..2523
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2805..3162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2805..2996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2828..2848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3013..3148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3177..3224
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3256..4050
                     /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(3310..3329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3920..3939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4114..4162
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4401..4989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4481..4500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5105..5388
                     /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 caccatgaac tccttctcca
     1081 caagcgcctt cggtccagtt gccttctccc tgggcctgct cctggtgttg cctgctgcct
     1141 tccctgcccc agtctccgtg agcggctggc ggctgttcaa gaagattagc gggagctccg
     1201 gtggctcgag cggtgcgatc gccatggagc tggcggtcgg gaacctcagc gagggcaacg
     1261 cgagctggcc ggagcccccc gccccggagc ccgggccgct gttcggcatc ggcgtggaga
     1321 acttcgtcac gctggtggtg ttcggcctga tcttcgcgct gggcgtgctg ggcaacagcc
     1381 tagtgatcac cgtgctggcg cgcagcaagc cgggcaagcc gcggagcacc accaacctgt
     1441 tcatcctcaa cctgagcatc gccgacctgg cctacctgct cttctgcatc cccttccagg
     1501 ccaccgtgta cgcgctgccc acctgggtgc tgggcgcctt catctgcaag ttcatccact
     1561 acttcttcac cgtgtccatg ctggtgagca tcttcaccct ggccgcgatg tccgtggacc
     1621 gctacgtggc catcgtgcac tcgcggcgct cctcctccct cagggtgtcc cgcaacgcgc
     1681 tgctgggcgt gggctgcatc tgggcgctgt ccattgccat ggcctcgccc gtggcctacc
     1741 accagggcct cttccacccg cgcgccagca accagacctt ctgctgggag cagtggcccg
     1801 accctcgcca caagaaggcc tacgtggtgt gcaccttcgt cttcggctac ctgctgccgc
     1861 tcctgctcat ctgcttctgc tatgccaagg tccttaatca cttgcataaa aagttgaaga
     1921 acatgtcaaa gaagtctgaa gcatccaaga aaaagactgc acagacagtt ctggtggtgg
     1981 ttgtggtgtt tggaatctcc tggctgccgc accacatcat ccatctctgg gctgagtttg
     2041 gagttttccc gctgacgccg gcttccttcc tcttcagaat caccgcccac tgcctggcgt
     2101 acagcaattc ctccgtgaat cctatcattt atgcatttct ctctgaaaat ttcaggaagg
     2161 cctataaaca agtgttcaag tgtcacattc gcaaagattc acacctgagt gatactaaag
     2221 aaaataaaag tcgaatagac accccaccat caaccaattg tactcatgtg gtttaaacga
     2281 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     2341 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     2401 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2461 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2521 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2581 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     2641 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     2701 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     2761 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     2821 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2881 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3001 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3061 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3121 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     3181 acaattaatc atcggcatag tatgtcggca tagtataata cgacaaggtg aggaactaaa
     3241 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3421 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3781 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4081 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     4141 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt
     4201 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     4261 ctggctttct acccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4321 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4381 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4441 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4501 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4561 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4621 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4681 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4741 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4801 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4861 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4921 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4981 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5041 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5101 tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt gaaaccatga
     5161 aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata
     5221 ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt
     5281 gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat
     5341 taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga taaaccgcag
     5401 tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
