LOCUS       Exported                6679 bp ds-DNA     circular SYN 02-JUN-2025
DEFINITION  Mammalian expression vector with the CMV promoter and a 
            neomycin-resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6679)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     The cloned gene must include a Kozak sequence and start codon.
FEATURES             Location/Qualifiers
     source          1..6679
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMVf
                     /note="Sequencing primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..942
                     /codon_start=1
                     /label=SmBiT
                     /translation="MVTGYRLFEEIL"
     CDS             943..975
                     /codon_start=1
                     /label=Linker
                     /translation="SGLKSRRALDS"
     CDS             976..2202
                     /codon_start=1
                     /label=New Arr3
                     /translation="MGEKPGTRVFKKSSPNCKLTVYLGKRDFVDHLDKVDPVDGVVLVD
                     PDYLKDRKVFVTLTCAFRYGREDLDVLGLSFRKDLFIANYQAFPPTPNPPRPPTRLQER
                     LLRKLGQHAHPFFFTIPQNLPCSVTLQPGPEDTGKACGVDFEIRAFCAKSLEEKSHKRN
                     SVRLVIRKVQFAPEKPGPQPSAETTRHFLMSDRSLHLEASLDKELYYHGEPLNVNVHVT
                     NNSTKTVKKIKVSVRQYADICLFSTAQYKCPVAQVEQDDQVSPSSTFCKVYTITPLLSN
                     NREKRGLALDGKLKHEDTNLASSTIVKEGANKEVLGILVSYRVKVKLVVSRGGDVSVEL
                     PFVLMHPKPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFEDFARLRLKGL
                     KDEDYDDQFC"
     CDS             976..1131
                     /codon_start=1
                     /label=T1
                     /translation="MGEKPGTRVFKKSSPNCKLTVYLGKRDFVDHLDKVDPVDGVVLVD
                     PDYLKDR"
     primer_bind     1068..1105
                     /label=AT2f
     CDS             1234..1302
                     /codon_start=1
                     /label=T2A
                     /translation="AFPPTPNPPRPPTRLQERLLRKL"
     CDS             1303..1347
                     /codon_start=1
                     /label=T2B G110-L124
                     /translation="GQHAHPFFFTIPQNL"
     CDS             1489..1599
                     /codon_start=1
                     /label=T3 V172-L208
                     /translation="VQFAPEKPGPQPSAETTRHFLMSDRSLHLEASLDKEL"
     CDS             1600..1653
                     /codon_start=1
                     /label=T4 Y209-T226
                     /translation="YYHGEPLNVNVHVTNNST"
     CDS             1771..1833
                     /codon_start=1
                     /label=T5 S226-R286
                     /translation="SSTFCKVYTITPLLSNNREKR"
     CDS             1834..1929
                     /codon_start=1
                     /label=T6 G287-I318
                     /translation="GLALDGKLKHEDTNLASSTIVKEGANKEVLGI"
     CDS             2008..2202
                     /codon_start=1
                     /label=T7 M345-C409
                     /translation="MHPKPHDHIALPRPQSAVPETDAPVDTNLIEFETNYATDDDIVFE
                     DFARLRLKGLKDEDYDDQFC"
     promoter        complement(2231..2249)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2275..2499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2369..2388)
                     /label=P3BHGR
                     /note="BGH Revese, sequencing primer"
     rep_origin      2545..2973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2987..3317
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3168..3303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3384..4178
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4352..4473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4522..4538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4924..5512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5683..6543)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6544..6648)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggaattc
      901 accaccatgg tgaccggcta ccggctgttc gaggagattc tctccggact taagtctcgg
      961 cgagcgctcg actccatggg ggagaaaccc gggaccaggg tcttcaagaa atcgagtcct
     1021 aactgcaagc tcaccgtgta cctaggcaag cgggactttg tggaccacct ggacaaagtg
     1081 gaccctgtag atggtgtggt gctagtggat ccggactact tgaaggaccg caaagtgttt
     1141 gtgaccctca cgtgcgcctt tcgctatggc cgcgaagacc tggacgtcct gggcttgtcc
     1201 ttccgcaaag acctgttcat cgccaactac caggccttcc ctccaacacc taacccacct
     1261 agaccaccta cgcgtctgca ggagcggctg ctgaggaagc tgggccagca tgcccacccc
     1321 ttttttttca caatacccca gaacctgccc tgctccgtca ccctacagcc aggcccagag
     1381 gatacgggga aggcttgcgg ggtcgacttt gagattcgag ccttctgcgc caaatcacta
     1441 gaagagaaaa gccacaaacg gaactccgta cgtctggtga tccgaaaggt gcagtttgcc
     1501 ccagagaaac cagggcccca gccttcagct gaaaccacac gccacttcct catgtctgac
     1561 cggtccctgc accttgaggc ttcactggac aaggagctgt actaccacgg ggagcccctc
     1621 aatgtcaatg tccacgtcac caataactcc accaagactg tcaagaagat caaagtctct
     1681 gtgagacagt acgccgacat ctgcctcttc agcaccgccc agtacaagtg ccccgtggct
     1741 caggtcgaac aagatgacca ggtgtcgccc agttccacgt tctgtaaggt gtacaccata
     1801 accccgctgc tcagcaacaa ccgggagaag cgcggcctcg ctctagatgg gaagctcaag
     1861 cacgaggaca ccaacctggc ctcgagcacc attgtgaagg agggtgccaa caaggaggtg
     1921 ctggggatac tggtgtccta cagggtcaag gtgaagctgg tggtgtcacg aggcggggat
     1981 gtctcagtgg agctgccatt tgttctcatg caccccaagc cccatgatca catcgccctt
     2041 cccaggcccc agtcagctgt tcctgaaaca gatgcccccg tggacaccaa cctcattgaa
     2101 tttgaaacca actatgccac agacgacgac atcgtgttcg aagactttgc ccggctgcgg
     2161 ctcaaggggc tgaaggacga ggactatgac gaccagttct gtatcgatta gggctggaag
     2221 cttgccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     2281 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2341 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2401 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2461 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2521 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2581 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2641 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2701 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2761 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2821 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2881 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     2941 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3001 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     3061 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3241 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3301 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3361 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3421 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3481 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3541 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3601 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3661 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3721 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3781 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3841 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3901 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3961 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4021 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4081 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4141 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4201 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4261 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4321 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4381 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4441 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4501 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4561 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4621 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4681 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5101 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5341 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6661 aaaagtgcca cctgacgtc
//
