LOCUS       sequence-405353-        4363 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Ready for cloning and heterologous expression of genes in mammalian 
            cells with protein export to cultivation media.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4363)
  AUTHORS   Meskova K, Martonova K, Hrasnova P, Sinska K, Skrabanova M, Fialova 
            L, Njemoga S, Cehlar O, Parmar O, Kolenko P, Pevala V, Skrabana R
  TITLE     Cost-Effective Protein Production in CHO Cells Following 
            Polyethylenimine-Mediated Gene Delivery Showcased by the Production 
            and Crystallization of Antibody Fabs.
  JOURNAL   Antibodies (Basel). 2023 Aug 4;12(3):51. doi: 10.3390/antib12030051.
  PUBMED    37606435
REFERENCE   2  (bases 1 to 4363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antibodies 
            (Basel)."; date: "2023-08-4"; volume: "12(3):51. doi"; pages: " 
            10.3390/antib12030051"
COMMENT     Sequence Label: pCMV-3UTR
FEATURES             Location/Qualifiers
     source          1..4363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1157..1278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1194..1213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1248..1267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1285..1740)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1422..1443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1634..1653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1767..1871
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1873..2230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1873..2064
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1896..1916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2081..2216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2265..3059
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2265..3059
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2319..2338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2929..2948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3247..3266)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3291..3338
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3667..4255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4156..4175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgccgccac catggaagcc ccagcgcagc ttctcttcct cctgctactc tggctccctg
      661 ataccaccgg tactagtgaa ttctcgagaa tattatccct aatacgtgcc accccactct
      721 taatcagtgg tggaagaacg gtctcagaac tgtttgtttc aattggccat ttaagtttag
      781 tagtaaaaga ctggttaatg ataacaatgc atcgtaaaac cttcagaagg aaaggagaat
      841 gttttgtgga ccactttggt tttctttttt gcgtgtggca gttttaagtt atcgatcgcc
      901 cttcccaaca gttgcgcagc ctgaatggcg aatggagatc caatttttaa gtgtataatg
      961 tgttaaacta ctgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt
     1021 caggcccctc agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga
     1081 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1141 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1201 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1261 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     1321 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1381 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1441 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1501 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1561 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1621 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1681 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1741 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1801 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1861 aaaggaagaa tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     1921 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1981 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2041 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2101 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2161 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2221 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2281 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2341 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2401 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     2461 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2521 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2581 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2641 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2701 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagaacat caggggctcg
     2761 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     2821 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     2881 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     2941 gtgatattgc tgaagaactt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3001 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3061 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3121 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3181 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     3241 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3301 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3361 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3421 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3481 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     3541 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3781 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcc
//
