LOCUS       sequence-481212-        5494 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express HiBiT-AVPR2 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 5494)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5494)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-AVPR2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5494
                     /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(2450..2468)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2505..2626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2516..2535)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2570..2589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2871..3228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2871..3062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2894..2914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3079..3214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3309..4103
                     /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(3363..3382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3973..3992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4167..4215
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4454..5042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4534..4553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5158..5441
                     /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 gccatgctca tggcgtccac cacttccgct gtgcctgggc
     1261 atccctctct gcccagcctg cccagcaaca gcagccagga gaggccactg gacacccggg
     1321 acccgctgct agcccgggcg gagctggcgc tgctctccat agtctttgtg gctgtggccc
     1381 tgagcaatgg cctggtgctg gcggccctag ctcggcgggg ccggcggggc cactgggcac
     1441 ccatacacgt cttcattggc cacttgtgcc tggccgacct ggccgtggct ctgttccaag
     1501 tgctgcccca gctggcctgg aaggccaccg accgcttccg tgggccagat gccctgtgtc
     1561 gggccgtgaa gtatctgcag atggtgggca tgtatgcctc ctcctacatg atcctggcca
     1621 tgacgctgga ccgccaccgt gccatctgcc gtcccatgct ggcgtaccgc catggaagtg
     1681 gggctcactg gaaccggccg gtgctagtgg cttgggcctt ctcgctcctt ctcagcctgc
     1741 cccagctctt catcttcgcc cagcgcaacg tggaaggtgg cagcggggtc actgactgct
     1801 gggcctgctt tgcggagccc tggggccgtc gcacctatgt cacctggatt gccctgatgg
     1861 tgttcgtggc acctaccctg ggtatcgccg cctgccaggt gctcatcttc cgggagattc
     1921 atgccagtct ggtgccaggg ccatcagaga ggcctggggg gcgccgcagg ggacgccgga
     1981 caggcagccc cggtgaggga gcccacgtgt cagcagctgt ggccaagact gtgaggatga
     2041 cgctagtgat tgtggtcgtc tatgtgctgt gctgggcacc cttcttcctg gtgcagctgt
     2101 gggccgcgtg ggacccggag gcacctctgg aaggggcgcc ctttgtgcta ctcatgttgc
     2161 tggccagcct caacagctgc accaacccct ggatctatgc atctttcagc agcagcgtgt
     2221 cctcagagct gcgaagcttg ctctgctgtg cccggggacg caccccaccc agcctgggtc
     2281 cccaagatga gtcctgcacc accgccagct cctccctggc caaggacact tcatcggttt
     2341 aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     2401 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2461 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2641 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     2701 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     2761 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     2821 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     2881 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa
     3241 ctgttgacaa ttaatcatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     3301 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4141 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     4201 ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg
     4261 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     4321 tctacccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4861 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     5161 gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg
     5221 cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc
     5281 atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt
     5341 ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga
     5401 ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg
     5461 gccacgatgc gtccggcgta gaggatcgag atct
//
