LOCUS       sequence-115794-        7045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  backbone for mouse recombinant antibody with added C-terminal FLAG 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7045)
  AUTHORS   Staudt N, Muller-Sienerth N, Fane-Dremucheva A, Yusaf SP, Millrine 
            D, Wright GJ
  TITLE     A panel of recombinant monoclonal antibodies against zebrafish 
            neural receptors and secreted proteins suitable for wholemount 
            immunostaining.
  JOURNAL   Biochem Biophys Res Commun. 2015 Jan 2;456(1):527-33. doi: 
            10.1016/j.bbrc.2014.11.123. Epub 2014 Dec 6.
  PUBMED    25490391
REFERENCE   2  (bases 1 to 7045)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbrc.2014.11.123"; journalName: "Biochem Biophys Res 
            Commun"; date: "2015-01-2- 2"; volume: "456"; issue: "1"; pages: 
            "527-33"
COMMENT     Sequence Label: pVk7-33G1-Flag
FEATURES             Location/Qualifiers
     source          1..7045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1239..1258
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1281..1290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2350..2373
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2425..2480
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2479..2498)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4716..4734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4802..4906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4907..4975,4976..5767)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5125..5144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5938..6526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6427..6446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6680..6697
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6814..6835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6850..6855,6856..6873,6874..6880)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6888..6904
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6893..6915
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6912..6928
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6912..6928
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
       61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
      781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
      841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
      901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
      961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     1261 ccactcccag gtccaagttt gccaccatgg agtttcagac ccaggtactc atgtccctgc
     1321 tgctctgcat gtctggtgcg gccgctccgg ctagcgataa gcttgacggc gcgcccgaca
     1381 cccccatctg tctatccact ggcccctgga tctgctgccc aaactaactc catggtgacc
     1441 ctgggatgcc tggtcaaggg ctatttccct gagccagtga cagtgacctg gaactctgga
     1501 tccctgtcca gcggtgtgca caccttccca gctgtcctgc agtctgacct ctacactctg
     1561 agcagctcag tgactgtccc ctccagcacc tggcccagcg agaccgtcac ctgcaacgtt
     1621 gcccacccgg ccagcagcac caaggtggac aagaaaattg tgcccaggga ttgtggttgt
     1681 aagccttgca tatgtacagt cccagaagta tcatctgtct tcatcttccc cccaaagccc
     1741 aaggatgtgc tcaccattac tctgactcct aaggtcacgt gtgttgtggt agacatcagc
     1801 aaggatgatc ccgaggtcca gttcagctgg tttgtagatg atgtggaggt gcacacagct
     1861 cagacgcaac cccgggagga gcagttcaac agcactttcc gctcagtcag tgaacttccc
     1921 atcatgcacc aggactggct caatggcaag gagttcaaat gcagggtcaa cagtgcagct
     1981 ttccctgccc ccatcgagaa aaccatctcc aaaaccaaag gcagaccgaa ggctccacag
     2041 gtgtacacca ttccacctcc caaggagcag atggccaagg ataaagtcag tctgacctgc
     2101 atgataacag acttcttccc tgaagacatt actgtggagt ggcagtggaa tgggcagcca
     2161 gcggagaact acaagaacac tcagcccatc atggacacag atggctctta cttcgtctac
     2221 agcaagctca atgtgcagaa gagcaactgg gaggcaggaa atactttcac ctgctctgtg
     2281 ttacatgagg gcctgcacaa ccaccatact gagaagagcc tctcccactc tcctggtaaa
     2341 tccggctccg attacaagga tgacgatgac aagtaaatct agacacgtga ttaattaagg
     2401 atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     2461 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga
     2521 tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta
     2581 cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca
     2641 acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt
     2701 ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct
     2761 ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta
     2821 actcttggct gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga
     2881 agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg
     2941 accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt
     3001 agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata
     3061 tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac
     3121 agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac
     3181 gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg
     3241 aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg
     3301 ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc
     3361 cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa
     3421 ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt
     3481 taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg
     3541 aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat
     3601 gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg
     3661 ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa
     3721 attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt
     3781 ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact
     3841 gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc
     3901 acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg
     3961 ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag
     4021 cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta
     4081 atgttgccat gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     4141 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     4201 atatctgggt agtatatgct atcctaattt atatctgggt agcataggct atcctaatct
     4261 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct
     4321 gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct atcctaattt
     4381 atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     4441 ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4501 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4561 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4621 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag
     4681 ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     4741 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4801 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4861 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4921 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4981 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5041 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5101 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     5161 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5221 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5281 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5341 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5401 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5461 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5521 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5581 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5641 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5701 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5761 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5821 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5881 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5941 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6001 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6061 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6121 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6181 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6241 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6301 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6361 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6421 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6481 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6541 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6601 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6661 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6721 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6781 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6841 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6901 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     6961 ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg
     7021 caggcgcaga actggtaggt atgga
//
