LOCUS       sequence-224250-        5998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-S tagged cytoplasmic catalytic domain 2 of Adenylate Cyclase 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5998)
  AUTHORS   Thomas A, Ramananda Y, Mun K, Naren AP, Arora K
  TITLE     AC6 is the major adenylate cyclase forming a diarrheagenic protein 
            complex with cystic fibrosis transmembrane conductance regulator in 
            cholera.
  JOURNAL   J Biol Chem. 2018 Jun 14. pii: RA118.003378. doi: 
            10.1074/jbc.RA118.003378.
  PUBMED    29903911
REFERENCE   2  (bases 1 to 5998)
  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"; journalName: "J Biol 
            Chem. 2018 Jun 14. pii: RA118.003378. doi: 
            10.1074/jbc.RA118.003378."
COMMENT     Sequence Label: pTriEx4-ADCY6-C2 (917-1165 aa)
FEATURES             Location/Qualifiers
     source          1..5998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..733
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     795..814
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     856..875
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    875..899
                     /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)."
     promoter        915..1024
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             1045..1062
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1072..1116
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1132..1149
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1168..1182
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2066..2098
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2105..2128
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2218..2237)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2283..2338
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2337..2356)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2426..2473
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2486..3191
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2574..2946
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2603..3191)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3200..3233)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3401..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3480..3499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4001..4792,4793..4861))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4624..4643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     5104..5931
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5104..5228
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5266..5871)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagacgtcg tttagtgaac cgtcagatca ctagatgctt
      541 tattgcggta gtttatcaca gttaaattgc taacgccagt ctcgaactta acgtgcagaa
      601 gttggtcgtg aggcactggg caggtaagta tcgggccctt tgtgcggggg gagcggctcg
      661 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
      721 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
      781 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
      841 ggatcggacc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      901 cggagttaat ccgggacctt taattcaacc caacacaata tattatagtt aaataagaat
      961 tattatcaaa tcatttgtat attaattaaa atactatact gtaaattaca ttttatttac
     1021 aatcaaagga gatataccat ggcacaccat caccaccatc actcttctgg taaagaaacc
     1081 gctgctgcga aatttgaacg ccagcacatg gactcgccac cgccttctgg tctggtcccc
     1141 cggggcagcg caggttctgg tacgattgat gacgacgaca agatggagtc gactgcccgc
     1201 ctagacttcc tctggaaact acaggcaaca ggggagaagg aggagatgga ggagctacag
     1261 gcatacaacc ggaggctgct gcataacatt ctgcccaagg acgtggcggc ccacttcctg
     1321 gcccgggagc gccgcaatga tgaactctac tatcagtcgt gtgagtgtgt ggctgttatg
     1381 tttgcctcca ttgccaactt ctctgagttc tatgtggagc tggaggcaaa caatgagggt
     1441 gccgagtgcc tgcggctgct caacgagatc atcgctgact ttgatgagat tatcagcgag
     1501 gagcggttcc ggcagctgga aaagatcaag acgattggta gcacctacat ggctgcctca
     1561 gggctgaacg ccagcaccta cgatcaggtg ggccgctccc acatcactgc cctggctgac
     1621 tacgccatgc ggctcatgga gcagatgaag cacatcaatg agcactcctt caacaatttc
     1681 cagatgaaga ttgggctgaa catgggccca gtcgtggcag gtgtcatcgg ggctcggaag
     1741 ccacagtatg acatctgggg gaacacagtg aatgtctcta gtcgtatgga cagcacgggg
     1801 gtccccgacc gaatccaggt gaccacggac ctgtaccagg ttctagctgc caagggctac
     1861 cagctggagt gtcgaggggt ggtcaaggtg aagggcaagg gggagatgac cacctacttc
     1921 ctcaatgggg gccccagcag ttaaaccggg cttctcctca acgatatctg agctcgtgga
     1981 tccgaattct cagatctcgg cgcgcctgca ggtcgacggt accggttcga agcttgcggc
     2041 cgcacagctg tatacacgtg caagccagcc agaactcgcc ccggaagacc ccgaggatct
     2101 cgagcaccac catcaccatc accatcacta agtgattaac ctcaggtgca ggctgcctat
     2161 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcgatct
     2221 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     2281 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2341 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     2401 tttggcaaca tatgcccata tgtaactagc ataacccctt ggggcctcta aacgggtctt
     2461 gaggggtttt ttgctgaaag catgcggagg aaattctcct tgaagtttcc ctggtgttca
     2521 aagtaaagga gtttgcacca gacgcacctc tgttcactgg tccggcgtat taaaacacga
     2581 tacattgtta ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca
     2641 attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag
     2701 cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag
     2761 atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg
     2821 gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc
     2881 tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa
     2941 tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa
     3001 acacgttaaa tagagcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat
     3061 tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag
     3121 ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc
     3181 tttttggaat tgcgatcgca taacttcgta tagcatacat tatacgaagt tataagctcg
     3241 gaacgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3301 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg ttaccaatgc ttaatcagtg
     4021 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4081 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4141 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4201 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4261 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4321 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4381 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4441 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4501 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4561 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4621 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4681 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4741 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4801 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4861 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgtccgcgc
     4921 gtttcctgca tcttttaatc aaatcccaag atgtgtataa acgcgccggt atgtacagga
     4981 agaggtttat actaaactgt tacattgcaa acgtggtttc gtgtgccaag tgtgaaaacc
     5041 gatgtttaat caaggctctg acgcatttct acaaccacga ctccaagtgt gtgggtgaag
     5101 tcatgcatct tttaatcaaa tcccaagatg tgtataaacc accaaactgc caaaaaatga
     5161 aaactgtcga caagctctgt ccgtttgctg gcaactgcaa gggtctcaat cctatttgta
     5221 attattgaat aataaaacaa ttataaatgt caaatttgtt ttttattaac gatacaaacc
     5281 aaacgcaaca agaacatttg tagtattatc tataattgaa aacgcgtagt tataatcgct
     5341 gaggtaatat ttaaaatcat tttcaaatga ttcacagtta atttgcgaca atataatttt
     5401 attttcacat aaactagacg ccttgtcgtc ttcttcttcg tattccttct ctttttcatt
     5461 tttctcttca taaaaattaa catagttatt atcgtatcca tatatgtatc tatcgtatag
     5521 agtaaatttt ttgttgtcat aaatatatat gtctttttta atggggtgta tagtaccgct
     5581 gcgcatagtt tttctgtaat ttacaacagt gctattttct ggtagttctt cggagtgtgt
     5641 tgctttaatt attaaattta tataatcaat gaatttggga tcgtcggttt tgtacaatat
     5701 gttgccggca tagtacgcag cttcttctag ttcaattaca ccatttttta gcagcaccgg
     5761 attaacataa ctttccaaaa tgttgtacga accgttaaac aaaaacagtt cacctccctt
     5821 ttctatacta ttgtctgcga gcagttgttt gttgttaaaa ataacagcca ttgtaatgag
     5881 acgcacaaac taatatcaca aactggaaat gtctatcaat atatagttgc tgatggccgg
     5941 cctattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
