LOCUS       Exported                7368 bp DNA     circular SYN 22-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  AUTHORS   RIKEN DNA Bank
  TITLE     Direct Submission
  JOURNAL   Exported Oct 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     RDB19691_pN1-cAMPinG1
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..1694
                     /label=CAG promoter
     enhancer        95..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..677
                     /label=chicken beta-actin promoter
     intron          678..1694
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             1803..3458
                     /codon_start=1
                     /label=cAMPinG1
                     /label=CyA033
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDEDAASYVR
                     KVIPKDYKTMAALAKAIEKNVLFSHLDDNERSDIFDAMFPVSFIAGETVIQQGDEGDNF
                     YVIDQGEMDVYVSWPVYIKADKQKNGIKANFKIWHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLLVQSSLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNGWATSVGEGGSFGELALIYGTPRAATVKAKTNVKLWGIDR
                     DSYRRILMGSTLRKRKMYEEFLSKVSILESLDKWERLTVADALEPVQFEDGQKIVVQGE
                     PGDEFFIILEGTAAVLQRRSENEEFVEVGRLGPSDYFGEIALLMNRPRAATVVARGPLK
                     CVKLDRPRFERVLGPCSDILKRNIQQYNSFVSLSV"
     CDS             1812..1829
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1833..1865
                     /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             1869..1892
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     misc_feature    3493..4081
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4110..4231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4331..4786)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4813..4917
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4919..5276
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4919..5110
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      5127..5262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5311..6105
                     /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"
     polyA_signal    6337..6384
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    6713..7301
                     /label=pUC ori
ORIGIN
        1 gcggccgcat gcattagtta ttaatgatgg aatctagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      421 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      481 aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg
      541 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      601 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      661 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      721 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      781 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      841 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
      961 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1021 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1081 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1141 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1201 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1261 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     1321 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1381 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1441 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1501 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1561 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1621 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1681 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     1741 aagaattgat ttgatacgta cggttgggca tcgctagggc tagactagta ccggtcgcca
     1801 ccatgggttc tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa
     1861 tgggtcggga tctgtacgac gatgacgata aggatctcgc aacaatggtc gacgaggatg
     1921 ctgcctccta cgttagaaag gttattccaa aagattataa gacaatggct gctttagcca
     1981 aggccatcga aaagaatgtg ctgttttcac accttgatga taacgagaga agtgacattt
     2041 ttgatgctat gtttccagtc tcctttattg ctggagagac ggttattcag caaggtgatg
     2101 aaggggataa cttctatgtg attgatcaag gagaaatgga tgtctatgtc agttggcctg
     2161 tctatatcaa ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atctggcaca
     2221 acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg
     2281 acggccccgt gctgctgccc gacaaccact acctgcttgt gcagtcctcg ctttcgaaag
     2341 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2401 ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg
     2461 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2521 acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga
     2581 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2641 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2701 agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca
     2761 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2821 ttaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg
     2881 gttgggcaac cagtgttggg gaaggaggga gctttggaga gctggctttg atttatggaa
     2941 cacccagagc agccactgtc aaagcaaaga caaacgtgaa actgtggggc atcgaccgag
     3001 acagctaccg aagaatcctc atgggaagca ctctgcgaaa gaggaagatg tatgaagaat
     3061 tccttagtaa agtgtctatt ttagagtctc tggacaagtg ggagcgtctc acagtagccg
     3121 atgcattgga gcctgtccag tttgaagatg gacagaagat cgtggtgcaa ggagagccgg
     3181 gggatgagtt cttcatcatt ttagagggca cagctgctgt gctgcagcgt cggtcagaaa
     3241 acgaagaatt tgttgaagtg ggacgactgg ggccttctga ttattttggt gaaattgccc
     3301 tgctgatgaa tcgtcctcgg gctgccactg tggttgcccg gggccctttg aagtgcgtta
     3361 agttggaccg gcctcggttt gaacgcgtcc ttggcccgtg ctcagacatc ctcaagcgga
     3421 acatccagca gtacaacagc ttcgtgtccc tgtccgtctg ataagcttga taggaattca
     3481 cgcgtactag ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3541 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3601 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3661 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3721 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3781 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3841 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3901 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3961 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4021 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4081 catcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4141 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4201 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4261 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4321 tattaattaa aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4381 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4441 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4501 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4561 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4621 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4681 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4741 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4801 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4861 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4921 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4981 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5041 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5101 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5161 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5221 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     5281 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5341 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5401 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5461 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     5521 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5581 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5641 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5701 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5761 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5821 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5881 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5941 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6001 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6061 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6121 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6181 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6241 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     6301 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     6361 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6421 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6481 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6541 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6601 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6841 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6901 ctgtatcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7081 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7201 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7261 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7321 ttttacggtt cctggccttt tgctggcctt ttgctcggat ccacatgt
//
