LOCUS       sequence-138293-        5698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His tagged protein for antibody production screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  AUTHORS   Khan S, Perry C, Tetreault ML, Henry D, Trimmer JS, Zimmerman AL, 
            Matthews G
  TITLE     A novel cyclic nucleotide-gated ion channel enriched in synaptic 
            terminals of isotocin neurons in zebrafish brain and pituitary.
  JOURNAL   Neuroscience. 2010 Jan 13;165(1):79-89. doi: 
            10.1016/j.neuroscience.2009.09.040. Epub 2009 Sep 22.
  PUBMED    19778592
REFERENCE   2  (bases 1 to 5698)
  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.neuroscience.2009.09.040"; journalName: "Neuroscience"; 
            date: "2010-01-13- 13"; volume: "165"; issue: "1"; pages: "79-89"
COMMENT     Sequence Label: pet28bZFCNGA5
FEATURES             Location/Qualifiers
     source          1..5698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(537..569)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(573..590)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(600..617)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             636..658
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    673..697
                     /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     complement(697..716)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(698..716)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(800..819)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1025..1102
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1103..2185
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1122..1141)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2994..3185
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3179..3201)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3287..3429
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3342..3361
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3444..3461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3615..4203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3695..3714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4314..4333)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4325..5140
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4325..5140
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4398..4417)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5232..5687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5369..5390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5581..5600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattccat gcttagcact taaaggtgaa tgttgacctc actgtttgta
      241 ccatccccat caatagtgct ttcattacca tcagaaagga atccactgcc agaatgattg
      301 gttctgcatt ccagagtggt aagtctctgc ttcaggtgct gctgagttgc tgtatattga
      361 gccaatagtc gagcaaatcg agtttgtaat atttcaatag tatcttccat cctctctact
      421 cttttctcaa tatcttccac ccctattccg tctgcctcca cctcctcgag caaaccctct
      481 tttataagaa tttgtcttcc cttctcttcc agaactttct tagcatctgg atcccgaccc
      541 atttgctgtc caccagtcat gctagccata tggctgccgc gcggcaccag gccgctgctg
      601 tgatgatgat gatgatggct gctgcccatg gtatatctcc ttcttaaagt taaacaaaat
      661 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
      721 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
      781 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
      841 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
      901 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
      961 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     1021 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     1081 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1141 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1201 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1261 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     1321 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     1381 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     1441 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     1501 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     1561 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     1621 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     1681 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     1741 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     1801 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     1861 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     1921 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     1981 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     2041 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     2101 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2161 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     2221 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2281 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2341 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2401 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     2461 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     2521 cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     2581 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     2641 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     2701 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     2761 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     2821 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     2881 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     2941 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     3001 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     3061 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     3121 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     3181 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3241 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     3301 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     3361 actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3421 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4021 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4261 taagggattt tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     4321 tgttatgagc catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat
     4381 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4441 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4501 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4561 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     4621 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     4681 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     4741 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     4801 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     4861 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     4921 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     4981 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     5041 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     5101 taaattgcag tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat
     5161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5221 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5281 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5341 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5401 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5461 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5521 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5581 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5641 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
