LOCUS       sequence-224252-        6343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-S tagged cytoplasmic catalytic domain 1 of Adenylate Cyclase 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  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 6343)
  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-C1(306-672 aa)
FEATURES             Location/Qualifiers
     source          1..6343
                     /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             complement(1757..1771)
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     CDS             2411..2443
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2450..2473
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2563..2582)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2628..2683
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2682..2701)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2771..2818
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2831..3536
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2919..3291
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2948..3536)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3545..3578)
                     /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(3746..4333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3825..3844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4346..5137,5138..5206))
                     /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     4969..4988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     5449..6276
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5449..5573
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5611..6216)
                     /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 agatgacaca ctatccagca
     1201 gaggtgtctc agcgccaggc ctttcaggag acccgcggtt acatccaggc ccggctccac
     1261 ctgcagcatg agaatcggca gcaggagcgg ctgctgctgt cggtattgcc ccagcacgtc
     1321 gccatggaga tgaaagaaga catcaacaca aaaaaagaag acatgatgtt ccacaagatc
     1381 tacatacaga agcatgacaa tgtcagcatc ctgtttgcag acattgaggg cttcaccagc
     1441 ctggcatccc agtgcactgc gcaggagctg gtcatgaccc tgaatgagct ctttgcccgg
     1501 tttgacaagc tggctgcgga gaatcactgc ctgaggatca agatcttggg ggactgttac
     1561 tactgtgtgt cagggctgcc ggaggcccgg gccgaccatg cccactgctg tgtggagatg
     1621 ggggtagaca tgattgaggc catctcgctg gtacgtgagg tgacaggtgt gaatgtgaac
     1681 atgcgcgtgg gcatccacag cgggcgcgtg cactgcggcg tccttggctt gcggaaatgg
     1741 cagttcgatg tgtggtccaa tgatgtgacc ctggccaacc acatggaggc aggaggccgg
     1801 gctggccgca tccacatcac tcgggcaaca ctgcagtacc tgaacgggga ctacgaggtg
     1861 gagccaggcc gtggtggcga gcgcaacgcg tacctcaagg agcagcacat tgagactttc
     1921 ctcatcctgg gcgccagcca gaaacggaaa gaggagaagg ccatgctggc caagctgcag
     1981 cggactcggg ccaactccat ggaagggctg atgccgcgct gggttcctga tcgtgccttc
     2041 tcccggacca aggactccaa ggccttccgc cagatgggca ttgatgattc cagcaaagac
     2101 aaccggggca cccaagatgc cctgaaccct gaggatgagg tggatgagtt cctgagccgt
     2161 gccatcgatg cccgcagcat tgatcagctg cggaaggacc atgtgcgccg gtttctgctc
     2221 accttccaga gagaggatct tgagaagaag tactcccgga aggtggatcc ccgcttcgga
     2281 gcctactaaa ccgggcttct cctcaacgat atctgagctc gtggatccga attctcagat
     2341 ctcggcgcgc ctgcaggtcg acggtaccgg ttcgaagctt gcggccgcac agctgtatac
     2401 acgtgcaagc cagccagaac tcgccccgga agaccccgag gatctcgagc accaccatca
     2461 ccatcaccat cactaagtga ttaacctcag gtgcaggctg cctatcagaa ggtggtggct
     2521 ggtgtggcca atgccctggc tcacaaatac cactgagatc gatctttttc cctctgccaa
     2581 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     2641 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2701 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     2761 ccatatgtaa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2821 gaaagcatgc ggaggaaatt ctccttgaag tttccctggt gttcaaagta aaggagtttg
     2881 caccagacgc acctctgttc actggtccgg cgtattaaaa cacgatacat tgttattagt
     2941 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     3001 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     3061 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     3121 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     3181 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     3241 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     3301 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaatagag
     3361 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     3421 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     3481 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt ggaattgcga
     3541 tcgcataact tcgtatagca tacattatac gaagttataa gctcggaacg ctgcgctcgg
     3601 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3661 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3721 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg ccccctgacg agcatcacaa
     3781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4321 aaaaggatct caagaagatc ctttgttacc aatgcttaat cagtgaggca cctatctcag
     4381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5221 aatattattg aagcatttat cagggttatt gtctcatgtc cgcgcgtttc ctgcatcttt
     5281 taatcaaatc ccaagatgtg tataaacgcg ccggtatgta caggaagagg tttatactaa
     5341 actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg
     5401 ctctgacgca tttctacaac cacgactcca agtgtgtggg tgaagtcatg catcttttaa
     5461 tcaaatccca agatgtgtat aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc
     5521 tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat ttgtaattat tgaataataa
     5581 aacaattata aatgtcaaat ttgtttttta ttaacgatac aaaccaaacg caacaagaac
     5641 atttgtagta ttatctataa ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa
     5701 atcattttca aatgattcac agttaatttg cgacaatata attttatttt cacataaact
     5761 agacgccttg tcgtcttctt cttcgtattc cttctctttt tcatttttct cttcataaaa
     5821 attaacatag ttattatcgt atccatatat gtatctatcg tatagagtaa attttttgtt
     5881 gtcataaata tatatgtctt ttttaatggg gtgtatagta ccgctgcgca tagtttttct
     5941 gtaatttaca acagtgctat tttctggtag ttcttcggag tgtgttgctt taattattaa
     6001 atttatataa tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc cggcatagta
     6061 cgcagcttct tctagttcaa ttacaccatt ttttagcagc accggattaa cataactttc
     6121 caaaatgttg tacgaaccgt taaacaaaaa cagttcacct cccttttcta tactattgtc
     6181 tgcgagcagt tgtttgttgt taaaaataac agccattgta atgagacgca caaactaata
     6241 tcacaaactg gaaatgtcta tcaatatata gttgctgatg gccggcctat taatagtaat
     6301 caattacggg gtcattagtt catagcccat atatggagtt ccg
//
