LOCUS       sequence-472506-        5412 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express AVPR2-SmBiT 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 5412)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AVPR2-SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    831..879
                     /note="synthetic polyadenylation signal"
     CDS             complement(943..1737)
                     /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(1054..1073)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1664..1683
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1769..1816)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1831..2188)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1845..1980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1997..2188
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2145..2165
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2433..2554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2470..2489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2524..2543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2591..2609
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(3912..3931)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3913..3931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3973..3992)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3975..4107
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4243..4446)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4272..4292)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4447..4826
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    5017..5300
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ttcttgaaat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgg
      721 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
      781 ctggacaagg gaaaacgcaa gcgcaaagag gatccttatc gctatcgatt cacacaaaaa
      841 accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt gatggcaggt
      901 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
      961 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     1021 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     1081 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     1141 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     1201 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     1261 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     1321 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     1381 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     1441 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     1501 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     1561 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     1621 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     1681 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgaa
     1741 ctgcctcctg ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg
     1801 atgattaatt gtcaacagtt aattaagctt tttgcaaaag cctaggcctc caaaaaagcc
     1861 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     1921 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     1981 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     2041 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     2101 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     2161 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     2221 tatttcaggc catggtgctg cgcagatccg cgtatgcggt gtgaaatacc gcacagatgc
     2281 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgatt
     2341 ttaccacatt tgtagaggtt ttacttgctt aaaaaaacct cccacatctc cccctgaacc
     2401 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2461 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2521 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccc
     2581 aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc
     2641 tttgttagca gccggatcag cttgcatgcc tgcaggtcga ctctagagtc gcggccttac
     2701 agaatctcct cgaacagccg gtagccggtc acaccgctcg agcctccacc tccgctcccg
     2761 ccaccacctg aactgccttg agaaaccgat gaagtgtcct tggccaggga ggagctggcg
     2821 gtggtgcagg actcatcttg gggacccagg ctgggtgggg tgcgtccccg ggcacagcag
     2881 agcaagcttc gcagctctga ggacacgctg ctgctgaaag atgcatagat ccaggggttg
     2941 gtgcagctgt tgaggctggc cagcaacatg agtagcacaa agggcgcccc ttccagaggt
     3001 gcctccgggt cccacgcggc ccacagctgc accaggaaga agggtgccca gcacagcaca
     3061 tagacgacca caatcactag cgtcatcctc acagtcttgg ccacagctgc tgacacgtgg
     3121 gctccctcac cggggctgcc tgtccggcgt cccctgcggc gccccccagg cctctctgat
     3181 ggccctggca ccagactggc atgaatctcc cggaagatga gcacctggca ggcggcgata
     3241 cccagggtag gtgccacgaa caccatcagg gcaatccagg tgacataggt gcgacggccc
     3301 cagggctccg caaagcaggc ccagcagtca gtgaccccgc tgccaccttc cacgttgcgc
     3361 tgggcgaaga tgaagagctg gggcaggctg agaaggagcg agaaggccca agccactagc
     3421 accggccggt tccagtgagc cccacttcca tggcggtacg ccaacatggg acggcagatg
     3481 gcacggtggc ggtccagcgt catggccagg atcatgtagg aggaggcata catgcccacc
     3541 atctgcagat acttcacggc ccgacacagg gcatctggcc cacggaagcg gtcggtggcc
     3601 ttccaggcca gctggggcag cacttggaac agagccacgg ccaggtcggc caggcacaag
     3661 tggccaatga agacgtgtat gggtgcccag tggccccgcc ggccccgccg agctagggcc
     3721 gccagcacca ggccattgct cagggccaca gccacaaaga ctatggagag cagcgccagc
     3781 tccgcccggg ctagcagcgg gtcccgggtg tccagtggcc tctcctggct gctgttgctg
     3841 ggcaggctgg gcagagaggg atgcccaggc acagcggaag tggtggacgc catgagcatg
     3901 gcgatcgcta gccctatagt gagtcgtatt aatactctag ccttaagagc tgtaattgaa
     3961 ctgggagtgg acacctgtgg agagaaaggc aaagtggatg tcagtaagac caataggtgc
     4021 ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc agtttctatt ggtctcctta
     4081 aacctgtctt gtaaccttga tacttacctg cccagtgcct cacgaccaac ttctgcagct
     4141 taagttcgag actgttgtgt cagaagcact gactgcgtta gcaatttaac tgtgataaac
     4201 taccgcaata aagcttctag tgatctgacg gttcactaaa ccagctctgc ttatatagac
     4261 ctcccaccgt acacgcctac cgcccatttg cgtcaacggg gcggggttat tacgacattt
     4321 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     4381 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttggtgtact gccaaaaccg
     4441 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtcccg
     4501 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     4561 gggcggactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     4621 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     4681 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     4741 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     4801 ttaataacta gtcaataatc aatgccaaca tggcggtcat attggacatg agccaatata
     4861 aatgtacata ttatgatata gatacaacgt atgcaatggc caatagccaa tattgattta
     4921 tgctatataa ccaatgaata atatggctaa tggccaatat tgaagatctc gatcctctac
     4981 gccggacgca tcgtggccgg agtactgcgg tttatctcga tggctacgag ggcagacagt
     5041 aagtggattt accataatcc cttaattgta cgcaccgcta aaacgcgttc agcgcgatca
     5101 cggcagcaga caggtaaaaa tggcaacaaa ccacccgaaa aactgccgca atcgcgcctg
     5161 ataaatttta accgtatgaa tacctatgca accagagggt acaggccaca ttacccccac
     5221 ttaatccact gaagctgcca tttttcatgg tttcaccatc ccagcgaagg gccatccagc
     5281 gtgcgttcct gtatttccgg actataaaag gatctaggtg aagatccttt ttgataatct
     5341 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5401 gatcaaagga tc
//
