LOCUS       sequence-466925-        5827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A-kinase anchor 
            protein 8 LCD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5827)
  TITLE     Mittag bacterial expression plasmids
REFERENCE   2  (bases 1 to 5827)
  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"
COMMENT     Sequence Label: pDest17_AKAP8 LCD (1-367)
FEATURES             Location/Qualifiers
     source          1..5827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1296..1320)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1387..1434
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1800..1805,1806..1822,1823..1828))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1855..1873)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1941..2045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2046..2114,2115..2906)
                     /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(2264..2283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3077..3665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3566..3585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3819..3836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3851..3990
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3916..3935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4076..4098
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4092..4283)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5745..5764
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggctccgaaa acctgtattt
      181 ccagggcagc atggatcagg gttatggtgg ttatggcgcc tggagtgccg gtccggccaa
      241 tacccagggt gcctatggca ccggcgttgc cagctggcag ggctatgaaa attataatta
      301 ttacggtgca cagaacacca gtgttaccac cggtgcaacc tatagctatg gcccggcaag
      361 ctgggaagcc gccaaagcaa atgatggtgg tctggccgca ggtgccccgg ctatgcatat
      421 ggcaagctat ggtccggaac cgtgcaccga taatagtgat agcctgattg caaaaattaa
      481 ccagcgcctg gatatgatga gcaaagaagg tggtcgtggc ggcagcggcg gtggtggtga
      541 aggtattcag gatcgtgaaa gcagttttcg ttttcagccg tttgaaagct atgatagccg
      601 cccgtgtctg ccggaacata atccgtatcg cccgagctat agctatgatt atgaatttga
      661 tctgggcagc gatcgtaatg gtagttttgg tggccagtat agcgaatgcc gtgatccggc
      721 ccgcgaacgt ggcagcctgg atggttttat gcgtggccgt ggccagggcc gttttcagga
      781 tcgcagtaat ccgggtacct ttatgcgcag cgatccgttt gtgccgccgg cagcaagtag
      841 cgaaccgctg agcaccccgt ggaatgaact gaattatgtt ggtggtcgcg gcctgggtgg
      901 cccgtcacct agtcgtccgc cgcctagcct gtttagtcag agcatggcac cggattatgg
      961 tgttatgggc atgcagggtg caggcggtta tgatagcacc atgccgtatg gctgcggtcg
     1021 tagtcagccg cgtatgcgcg atcgtgatcg tccgaaacgc cgcggctttg atcgctttgg
     1081 tccggatggt accggccgta aacgcaaaca gtttcagctg tatgaagaac cggataccaa
     1141 actggcccgc gttgatagtg aaggcgattt tagcgaaaat gatgatgcag caggcgattt
     1201 tcgtagtggc gatgaagaat tcaaaggcga agatgaactg tgcgatagtg gccgccagcg
     1261 cggtgaaaaa gaagatgaag atgaagacgt ttaaaaccca gctttcttgt acaaagtggt
     1321 tgattcgagg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1381 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1441 ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
     1501 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
     1561 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     1621 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat
     1681 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     1741 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     1801 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
     1861 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2281 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2581 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3961 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4021 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4081 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4141 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4201 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4261 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4321 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4381 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4441 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4501 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4561 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4621 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4681 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4741 cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     4801 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     4861 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     4921 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     4981 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     5041 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     5101 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     5161 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     5221 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     5281 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     5341 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     5401 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     5461 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     5521 tgaaggctct caagggcatc ggtcgatcga cgctctccct tatgcgactc ctgcattagg
     5581 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5641 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     5701 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     5761 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     5821 aggatcg
//
