LOCUS       sequence-478451-        5537 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express HiBiT-NPY2R 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 5537)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5537)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-NPY2R Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5537
                     /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(2480..2498)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2535..2656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2546..2565)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2600..2619
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2901..3258
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2901..3092
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2924..2944)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3109..3244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3273..3320
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3352..4146
                     /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(3406..3425)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4016..4035
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4210..4258
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4497..5085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4577..4596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5201..5484
                     /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 gccatgggtc caataggtgc agaggctgat gagaaccaga
     1261 cagtggaaga aatgaaggtg gaacaatacg ggccacaaac aactcctaga ggtgaactgg
     1321 tccctgaccc tgagccagag cttatagata gtaccaagct gattgaggta caagttgttc
     1381 tcatattggc ctactgctcc atcatcttgc ttggggtaat tggcaactcc ttggtgatcc
     1441 atgtggtgat caaattcaag agcatgcgca cagtaaccaa ctttttcatt gccaatctgg
     1501 ctgtggcaga tcttttggtg aacactctgt gtctaccgtt cactcttacc tataccttaa
     1561 tgggggagtg gaaaatgggt cctgtcctgt gccacctggt gccctatgcc cagggcctgg
     1621 cagtacaagt atccacaatc accttgacag taattgccct ggaccggcac aggtgcatcg
     1681 tctaccacct agagagcaag atctccaagc gaatcagctt cctgattatt ggcttggcct
     1741 ggggcatcag tgccctgctg gcaagtcccc tggccatctt ccgggagtat tcgctgattg
     1801 agatcatccc ggactttgag attgtggcct gtactgaaaa gtggcctggc gaggagaaga
     1861 gcatctatgg cactgtctat agtctttctt ccttgttgat cttgtatgtt ttgcctctgg
     1921 gcattatatc attttcctac actcgcattt ggagtaaatt gaagaaccat gtcagtcctg
     1981 gagctgcaaa tgaccactac catcagcgaa ggcaaaaaac caccaaaatg ctggtgtgtg
     2041 tggtggtggt gtttgcggtc agctggctgc ctctccatgc cttccagctt gccgttgaca
     2101 ttgacagcca ggtcctggac ctgaaggagt acaaactcat cttcacagtg ttccacatta
     2161 tcgccatgtg ctccactttt gccaatcccc ttctctatgg ctggatgaac agcaactaca
     2221 gaaaggcttt cctctcggcc ttccgctgtg agcagcggtt ggatgccatt cactctgagg
     2281 tgtccgtgac attcaaggct aaaaagaacc tggaggtcag aaagaacagt ggccccaatg
     2341 actctttcac agaggctacc aatgtcgttt aaacgaattc gggctcggta cccggggatc
     2401 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     2461 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     2521 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2581 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2641 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2701 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     2761 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     2821 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     2881 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     2941 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3001 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3061 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3121 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3181 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3241 ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata
     3301 tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa
     3361 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3421 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3481 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3541 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3601 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3661 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3721 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3781 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3841 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3901 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3961 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4021 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4081 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4141 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4201 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     4261 tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     4321 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctaccc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac
     5221 gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa
     5281 gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat
     5341 caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg
     5401 tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt
     5461 actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc
     5521 gtagaggatc gagatct
//
