LOCUS       sequence-224293-        8200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal Cyan Fluorescent tag in Adenylate Cyclase 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8200)
  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 8200)
  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: pAmCyan1-C1-ADCY6
FEATURES             Location/Qualifiers
     source          1..8200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1239
                     /codon_start=1
                     /product="Anemonia majano cyan fluorescent protein"
                     /label=AmCyan1
                     /note="mammalian codon-optimized"
                     /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST
                     FKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG
                     GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKKTTGWDPSFEKMTVCDGILKGDV
                     TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHVVEHRIARTDLDKGGNSVQLTEHAVAH
                     ITSVVPF"
     CDS             complement(2747..2761)
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     polyA_signal    4928..5049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4965..4984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5019..5038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5056..5511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5193..5214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5405..5424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5538..5642
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5644..6001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5644..5835
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5667..5687)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5852..5987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6036..6830
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6036..6830
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6090..6109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6700..6719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7018..7037)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7062..7109
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7438..8026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7927..7946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggccct gtccaacaag ttcatcggcg acgacatgaa gatgacctac
      601 cacatggacg gctgcgtgaa cggccactac ttcaccgtga agggcgaggg cagcggcaag
      661 ccctacgagg gcacccagac ctccaccttc aaggtgacca tggccaacgg cggccccctg
      721 gccttctcct tcgacatcct gtccaccgtg ttcatgtacg gcaaccgctg cttcaccgcc
      781 taccccacca gcatgcccga ctacttcaag caggccttcc ccgacggcat gtcctacgag
      841 agaaccttca cctacgagga cggcggcgtg gccaccgcca gctgggagat cagcctgaag
      901 ggcaactgct tcgagcacaa gtccaccttc cacggcgtga acttccccgc cgacggcccc
      961 gtgatggcca agaagaccac cggctgggac ccctccttcg agaagatgac cgtgtgcgac
     1021 ggcatcttga agggcgacgt gaccgccttc ctgatgctgc agggcggcgg caactacaga
     1081 tgccagttcc acacctccta caagaccaag aagcccgtga ccatgccccc caaccacgtg
     1141 gtggagcacc gcatcgccag aaccgacctg gacaagggcg gcaacagcgt gcagctgacc
     1201 gagcacgccg tggcccacat cacctccgtg gtgcccttct ccggactcag atctcgagct
     1261 atgtcatggt ttagtggcct cctggtccct aaagtggatg aacggaaaac agcctggggt
     1321 gaacgcaatg ggcagaagcg ttcgcggcgc cgtggcactc gggcaggtgg cttctgcacg
     1381 ccccgctata tgagctgcct ccgggatgca gagccaccca gccccacccc tgcgggcccc
     1441 cctcggtgcc cctggcagga tgacgccttc atccggaggg gcggcccagg caagggcaag
     1501 gagctggggc tgcgggcagt ggccctgggc ttcgaggata ccgaggtgac aacgacagcg
     1561 ggcgggacgg ctgaggtggc gcccgacgcg gtgcccagga gtgggcgatc ctgctggcgc
     1621 cgtctggtgc aggtgttcca gtcgaagcag ttccgttcgg ccaagctgga gcgcctgtac
     1681 cagcggtact tcttccagat gaaccagagc agcctgacgc tgctgatggc ggtgctggtg
     1741 ctgctcacag cggtgctgct ggctttccac gccgcacccg cccgccctca gcctgcctat
     1801 gtggcactgt tggcctgtgc cgccgccctg ttcgtggggc tcatggtggt gtgtaaccgg
     1861 catagcttcc gccaggactc catgtgggtg gtgagttacg tggtgctggg catcctggcg
     1921 gcagtgcagg tcgggggcgc tctcgcagca gacccgcgca gcccctctgc gggcctctgg
     1981 tgccctgtgt tctttgtcta catcgcctac acgctcctcc ccatccgcat gcgggctgcc
     2041 gtcctcagcg gcctgggcct ctccaccttg catttgatct tggcctggca acttaaccgt
     2101 ggtgatgcct tcctctggaa gcagctcggt gccaatgtgc tgctgttcct ctgcaccaac
     2161 gtcattggca tctgcacaca ctatccagca gaggtgtctc agcgccaggc ctttcaggag
     2221 acccgcggtt acatccaggc ccggctccac ctgcagcatg agaatcggca gcaggagcgg
     2281 ctgctgctgt cggtattgcc ccagcacgtt gccatggaga tgaaagaaga catcaacaca
     2341 aaaaaagaag acatgatgtt ccacaagatc tacatacaga agcatgacaa tgtcagcatc
     2401 ctgtttgcag acattgaggg cttcaccagc ctggcatccc agtgcactgc gcaggagctg
     2461 gtcatgaccc tgaatgagct ctttgcccgg tttgacaagc tggctgcgga gaatcactgc
     2521 ctgaggatca agatcttggg ggactgttac tactgtgtgt cagggctgcc ggaggcccgg
     2581 gccgaccatg cccactgctg tgtggagatg ggggtagaca tgattgaggc catctcgctg
     2641 gtacgtgagg tgacaggtgt gaatgtgaac atgcgcgtgg gcatccacag cgggcgcgtg
     2701 cactgcggcg tccttggctt gcggaaatgg cagttcgatg tgtggtccaa tgatgtgacc
     2761 ctggccaacc acatggaggc aggaggccgg gctggccgca tccacatcac tcgggcaaca
     2821 ctgcagtacc tgaacgggga ctacgaggtg gagccaggcc gtggtggcga gcgcaacgcg
     2881 tacctcaagg agcagcacat tgagactttc ctcatcctgg gcgccagcca gaaacggaaa
     2941 gaggagaagg ccatgctggc caagctgcag cggactcggg ccaactccat ggaagggctg
     3001 atgccgcgct gggttcctga tcgtgccttc tcccggacca aggactccaa ggccttccgc
     3061 cagatgggca ttgatgattc cagcaaagac aaccggggca cccaagatgc cctgaaccct
     3121 gaggatgagg tggatgagtt cctgagccgt gccatcgatg cccgcagcat tgatcagctg
     3181 cggaaggacc atgtgcgccg gtttctgctc accttccaga gagaggatct tgagaagaag
     3241 tactcccgga aggtggatcc ccgcttcgga gcctacgttg cctgtgccct gttggtcttc
     3301 tgcttcatct gcttcatcca gcttctcatc ttcccacact ccaccctgat gcttgggatc
     3361 tatgccagca tcttcctgct gctgctaatc accgtgctga tctgtgctgt gtactcctgt
     3421 ggttctctgt tccctaaggc cctgcaacgt ctgtcccgca gcattgtccg ctcacgggca
     3481 catagcaccg cagttggcat cttttccgtc ctgcttgtgt ttacttctgc cattgccaac
     3541 atgttcacct gtaaccacac ccccatacgg agctgtgcag cccggatgct gaatttaaca
     3601 cctgctgaca tcactgcctg ccacctgcag cagctcaatt actctctggg cctggatgct
     3661 cccctgtgtg agggcaccat gcccacctgc agctttcctg agtacttcat cgggaacatg
     3721 ctgctgagtc tcttggccag ctctgtcttc ctgcacatca gcagcatcgg gaagttggcc
     3781 atgatctttg tcttggggct catctatttg gtgctgcttc tgctgggtcc cccagccacc
     3841 atctttgaca actatgacct actgcttggc gtccatggct tggcttcttc caatgagacc
     3901 tttgatgggc tggactgtcc agctgcaggg agggtggccc tcaaatatat gacccctgtg
     3961 attctgctgg tgtttgcgct ggcgctgtat ctgcatgctc agcaggtgga gtcgactgcc
     4021 cgcctagact tcctctggaa actacaggca acaggggaga aggaggagat ggaggagcta
     4081 caggcataca accggaggct gctgcataac attctgccca aggacgtggc ggcccacttc
     4141 ctggcccggg agcgccgcaa tgatgaactc tactatcagt cgtgtgagtg tgtggctgtt
     4201 atgtttgcct ccattgccaa cttctctgag ttctatgtgg agctggaggc aaacaatgag
     4261 ggtgtcgagt gcctgcggct gctcaacgag atcatcgctg actttgatga gattatcagc
     4321 gaggagcggt tccggcagct ggaaaagatc aagacgattg gtagcaccta catggctgcc
     4381 tcagggctga acgccagcac ctacgatcag gtgggccgct cccacatcac tgccctggct
     4441 gactacgcca tgcggctcat ggagcagatg aagcacatca atgagcactc cttcaacaat
     4501 ttccagatga agattgggct gaacatgggc ccagtcgtgg caggtgtcat cggggctcgg
     4561 aagccacagt atgacatctg ggggaacaca gtgaatgtct ctagtcgtat ggacagcacg
     4621 ggggtccccg accgaatcca ggtgaccacg gacctgtacc aggttctagc tgccaagggc
     4681 taccagctgg agtgtcgagg ggtggtcaag gtgaagggca agggggagat gaccacctac
     4741 ttcctcaatg ggggccccag cagttaagaa ttctgcagtc gacggtaccg cgggcccggg
     4801 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     4861 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4921 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4981 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5041 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5101 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5161 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5221 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5281 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5341 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5401 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5461 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     5521 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5581 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5641 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5701 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5761 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5821 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5881 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5941 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6001 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6061 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6121 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6181 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     6241 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6301 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6361 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6421 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6481 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6541 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     6601 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6661 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6721 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6781 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6841 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6901 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6961 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     7021 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     7081 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     7141 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     7201 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     7261 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     7321 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7381 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7441 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7501 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7561 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7621 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7681 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7741 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7801 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7861 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7921 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7981 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8041 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     8101 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     8161 ttacggggtc attagttcat agcccatata tggagttccg
//
