LOCUS       sequence-115793-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  backbone for mouse recombinant antibody with added C-terminal 
            biotinylation site, linker, 6His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  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 7132)
  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-BioLHis
FEATURES             Location/Qualifiers
     source          1..7132
                     /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             2443..2460
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2512..2567
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2566..2585)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4803..4821)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4889..4993
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4994..5062,5063..5854)
                     /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(5212..5231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6025..6613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6514..6533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6767..6784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6901..6922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6937..6942,6943..6960,6961..6967)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6975..6991
                     /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     6980..7002
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6999..7015
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6999..7015
                     /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 tccggctcca attccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac
     2401 cacagggaca ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat
     2461 tgagtctaga cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga
     2521 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     2581 atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc
     2641 ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc
     2701 atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca
     2761 cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat
     2821 gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac
     2881 tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg
     2941 ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga
     3001 ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata
     3061 aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat
     3121 ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt
     3181 agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt
     3241 tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag
     3301 tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc
     3361 tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg
     3421 ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg
     3481 tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt
     3541 gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc
     3601 gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt
     3661 gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca
     3721 tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc
     3781 cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata
     3841 aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc
     3901 ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt
     3961 aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg
     4021 gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag
     4081 gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca
     4141 cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata
     4201 tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata
     4261 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata
     4321 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4381 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga
     4441 ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg
     4501 gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4561 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4621 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     4681 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     4741 ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa
     4801 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4861 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4921 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4981 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5041 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5101 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5161 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5221 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     5281 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5341 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5401 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5461 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5521 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5581 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5641 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5701 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5761 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5821 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5881 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5941 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6001 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6061 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6721 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6781 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6841 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6901 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6961 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     7021 ttacgccaag ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag
     7081 atccgggcaa cgttgttgcc attgctgcag gcgcagaact ggtaggtatg ga
//
