LOCUS       sequence-478487-        5735 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-ADRA2B 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 5735)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5735)
  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            "Unpublished"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-ADRA2B Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5735
                     /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(2678..2696)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2733..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3099..3456
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3099..3290
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3122..3142)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3307..3442
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3471..3518
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3550..4344
                     /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(3604..3623)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4214..4233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4408..4456
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4695..5283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4775..4794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5399..5682
                     /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 gccatggacc accaggaccc ctactccgtg caggccacag
     1261 cggccatagc ggcggccatc accttcctca ttctctttac catcttcggc aacgctctgg
     1321 tcatcctggc tgtgttgacc agccgctcgc tgcgcgcccc tcagaacctg ttcctggtgt
     1381 cgctggccgc cgccgacatc ctggtggcca cgctcatcat ccctttctcg ctggccaacg
     1441 agctgctggg ctactggtac ttccggcgca cgtggtgcga ggtgtacctg gcgctcgacg
     1501 tgctcttctg cacctcgtcc atcgtgcacc tgtgcgccat cagcctggac cgctactggg
     1561 ccgtgagccg cgcgctggag tacaactcca agcgcacccc gcgccgcatc aagtgcatca
     1621 tcctcactgt gtggctcatc gccgccgtca tctcgctgcc gcccctcatc tacaagggcg
     1681 accagggccc ccagccgcgc gggcgccccc agtgcaagct caaccaggag gcctggtaca
     1741 tcctggcctc cagcatcgga tctttctttg ctccttgcct catcatgatc cttgtctacc
     1801 tgcgcatcta cctgatcgcc aaacgcagca accgcagagg tcccagggcc aagggggggc
     1861 ctgggcaggg tgagtccaag cagccccgac ccgaccatgg tggggctttg gcctcagcca
     1921 aactgccagc cctggcctct gtggcttctg ccagagaggt caacggacac tcgaagtcca
     1981 ctggggagaa ggaggagggg gagacccctg aagatactgg gacccgggcc ttgccaccca
     2041 gttgggctgc ccttcccaac tcaggccagg gccagaagga gggtgtttgt ggggcatctc
     2101 cagaggatga agctgaagag gaggaggagg aggaggaaga gtgtgaaccc caggcagtgc
     2161 cagtgtctcc ggcctcagct tgcagccccc cgctgcagca gccacagggc tcccgggtgc
     2221 tggccaccct acgtggccag gtgctcctgg gcaggggcgt gggtgctata ggtgggcagt
     2281 ggtggcgtcg acgggcgcag ctgacccggg agaagcgctt caccttcgtg ctggctgtgg
     2341 tcattggcgt ttttgtgctc tgctggttcc ccttcttctt cagctacagc ctgggagcca
     2401 tctgcccgaa gcactgcaag gtgccccatg gcctcttcca gttcttcttc tggatcggct
     2461 actgcaacag ctcactgaac cctgttatct acaccatctt caaccaggac ttccgccgtg
     2521 ccttccggag gatcctgtgc cgcccgtgga cccagacggc ctgggtttaa acgaattcgg
     2581 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagctga tccggctgct
     2641 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2701 ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     2941 tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     3001 ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct
     3061 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     3121 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3181 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3301 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3361 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3421 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact gttgacaatt
     3481 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     3541 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3601 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3661 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3721 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3781 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3841 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3901 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3961 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4021 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4081 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4141 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4201 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4261 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4321 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4381 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     4441 gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt
     4501 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     4561 ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4861 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5101 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt
     5281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc
     5401 ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg
     5461 gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt
     5521 catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt
     5581 tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg
     5641 attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc
     5701 ggccacgatg cgtccggcgt agaggatcga gatct
//
