LOCUS       sequence-478486-        5780 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-ADRA2C 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 5780)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5780)
  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            "Unpublished"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: HiBiT-ADRA2C Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5780
                     /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(2723..2741)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2778..2899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2843..2862
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3144..3501
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3144..3335
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3167..3187)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3352..3487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3516..3563
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3595..4389
                     /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(3649..3668)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4259..4278
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4453..4501
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4740..5328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4820..4839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5444..5727
                     /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 gccatggcgt ccccggcgct ggcggcggcg ctggcggtgg
     1261 cggcagcggc gggccccaat gcgagcggcg cgggcgagag gggcagcggc ggggttgcca
     1321 atgcctcggg ggcttcctgg gggccgccgc gcggccagta ctcggcgggc gcggtggcag
     1381 ggctggctgc cgtggtgggc ttcctcatcg tcttcaccgt ggtgggcaac gtgctggtgg
     1441 tgatcgccgt gctgaccagc cgggcgctgc gcgcgccaca gaacctcttc ctggtgtcgc
     1501 tggcctcggc cgacatcctg gtggccacgc tggtcatgcc cttctcgttg gccaacgagc
     1561 tcatggccta ctggtacttc gggcaggtgt ggtgcggcgt gtacctggcg ctcgatgtgc
     1621 tgttttgcac ctcgtcgatc gtgcatctgt gtgccatcag cctggaccgc tactggtcgg
     1681 tgacgcaggc cgtcgagtac aacctgaagc gcacaccacg ccgcgtcaag gccaccatcg
     1741 tggccgtgtg gctcatctcg gccgtcatct ccttcccgcc gctggtctcg ctctaccgcc
     1801 agcccgacgg cgccgcctac ccgcagtgcg gcctcaacga cgagacctgg tacatcctgt
     1861 cctcctgcat cggctccttc ttcgcgccct gcctcatcat gggcctggtc tacgcgcgca
     1921 tctaccgagt ggccaagctg cgcacgcgca cgctcagcga gaagcgcgcc cccgtgggcc
     1981 ccgacggtgc gtccccgact accgaaaacg ggctgggcgc ggcggcaggc gcaggcgaga
     2041 acgggcactg cgcgcccccg cccgccgacg tggagccgga cgagagcagc gcagcggccg
     2101 agaggcggcg gcgccggggc gcgttgcggc ggggcgggcg gcggcgagcg ggcgcggagg
     2161 ggggcgcggg cggtgcggac gggcaggggg cggggccggg ggcggctgag tcgggggcac
     2221 tgaccgcctc caggtccccg gggcccggtg gccgcctgtc gcgcgccagc tcgcgctccg
     2281 tcgagttctt cctgtcgcgc cggcgccggg cgcgcagcag cgtgtgccgc cgcaaggtgg
     2341 cccaggcgcg cgagaagcgc ttcacctttg tgctggctgt ggtcatgggc gtgttcgtgc
     2401 tctgctggtt ccccttcttc ttcagctaca gcctgtacgg catctgccgc gaggcctgcc
     2461 aggtgcccgg cccgctcttc aagttcttct tctggatcgg ctactgcaac agctcgctca
     2521 acccggtcat ctacacggtc ttcaaccagg atttccggcg atcctttaag cacatcctct
     2581 tccgacggag gagaaggggc ttcaggcagg tttaaacgaa ttcgggctcg gtacccgggg
     2641 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     2701 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     2761 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2821 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2881 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2941 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3001 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3121 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3181 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3241 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3361 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3421 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3481 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     3541 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     3601 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3661 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3721 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3781 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3841 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3901 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3961 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4021 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4081 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4141 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4201 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4261 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4321 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4381 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4441 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4501 gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag
     4561 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cccggtatca
     4621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5161 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     5341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5401 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     5461 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     5521 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     5581 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     5641 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     5701 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     5761 ggcgtagagg atcgagatct
//
