LOCUS       sequence-478452-        5513 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-P2RY1 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 5513)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5513)
  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            "Unpublished"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-P2RY1 Vector
FEATURES             Location/Qualifiers
     source          1..5513
                     /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(2456..2474)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2511..2632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2522..2541)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2576..2595
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2877..3234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2877..3068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2900..2920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3085..3220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3249..3296
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3328..4122
                     /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(3382..3401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3992..4011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4186..4234
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4473..5061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4553..4572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5177..5460
                     /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 gccatgaccg aggtgctgtg gccggctgtc cccaacggga
     1261 cggacgctgc cttcctggcc ggtccgggtt cgtcctgggg gaacagcacg gtcgcctcca
     1321 ctgccgccgt ctcctcgtcg ttcaaatgcg ccttgaccaa gacgggcttc cagttttact
     1381 acctgccggc tgtctacatc ttggtattca tcatcggctt cctgggcaac agcgtggcca
     1441 tctggatgtt cgtcttccac atgaagccct ggagcggcat ctccgtgtac atgttcaatt
     1501 tggctctggc cgacttcttg tacgtgctga ctctgccagc cctgatcttc tactacttca
     1561 ataaaacaga ctggatcttc ggggatgcca tgtgtaaact gcagaggttc atctttcatg
     1621 tgaacctcta tggcagcatc ttgtttctga catgcatcag tgcccaccgg tacagcggtg
     1681 tggtgtaccc cctcaagtcc ctgggccggc tcaaaaagaa gaatgcgatc tgtatcagcg
     1741 tgctggtgtg gctcattgtg gtggtggcga tctcccccat cctcttctac tcaggtaccg
     1801 gggtccgcaa aaacaaaacc atcacctgtt acgacaccac ctcagacgag tacctgcgaa
     1861 gttatttcat ctacagcatg tgcacgaccg tggccatgtt ctgtgtcccc ttggtgctga
     1921 ttctgggctg ttacggatta attgtgagag ctttgattta caaagatctg gacaactctc
     1981 ctctgaggag aaaatcgatt tacctggtaa tcattgtact gactgttttt gctgtgtctt
     2041 acatcccttt ccatgtgatg aaaacgatga acttgagggc ccggcttgat tttcagaccc
     2101 cagcaatgtg tgctttcaat gacagggttt atgccacgta tcaggtgaca agaggtctag
     2161 caagtctcaa cagttgtgtg gaccccattc tctatttctt ggcgggagat actttcagaa
     2221 ggagactctc ccgagccaca aggaaagctt ctagaagaag tgaggcaaat ttgcaatcca
     2281 agagtgaaga catgaccctc aatattttac ctgagttcaa gcagaatgga gatacaagcc
     2341 tggtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
     2401 caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2461 gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga taagatacat
     2521 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2581 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2641 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     2701 gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg cttacaattt
     2761 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     2821 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     2881 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3061 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     3241 aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3301 tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct
     3361 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3421 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3481 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3541 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3601 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3661 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3721 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3781 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3841 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3901 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3961 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4021 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4081 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4141 aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta
     4201 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatcctcttt
     4261 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     4321 cgtttctgcg gactggcttt ctacccggta tcagctcact caaaggcggt aatacggtta
     4381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5041 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5161 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg
     5221 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc
     5281 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc
     5341 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc
     5401 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga
     5461 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
