LOCUS       sequence-14182-e        7170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7170)
  AUTHORS   Adalat S, Woolf AS, Johnstone KA, Wirsing A, Harries LW, Long DA, 
            Hennekam RC, Ledermann SE, Rees L, van't Hoff W, Marks SD, Trompeter
            RS, Tullus K, Winyard PJ, Cansick J, Mushtaq I, Dhillon HK, Bingham 
            C, Edghill EL, Shroff R, Stanescu H, Ryffel GU, Ellard S, 
            Bockenhauer D
  TITLE     HNF1B mutations associate with hypomagnesemia and renal magnesium 
            wasting.
  JOURNAL   J Am Soc Nephrol. 2009 May . 20(5):1123-31.
  PUBMED    19389850
REFERENCE   2  (bases 1 to 7170)
  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 Am Soc 
            Nephrol. 2009 May . 20(5):1123-31."
COMMENT     Sequence Label: FXYD2-luc
FEATURES             Location/Qualifiers
     source          1..7170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1938..1947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2434..2443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2440..4092
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2506..2524)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4133..4254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4144..4163)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4198..4217
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4502..4519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4673..5261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4753..4772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5432..6223,6224..6292))
                     /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     6055..6074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6293..6397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6424..6879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6511..6530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6721..6742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7010..7058
                     /note="synthetic polyadenylation signal"
     misc_feature    7072..7163
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7112..7131
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacctctg ttggtgcctc cccatgcacc tcctcccccc cgcagtggga gcccttccat
       61 ccacactgca gctttgtttg tgaccagggt aaatattacc tggcactgtt aatgtttaat
      121 gcccccacct catccccagc caccccacca aggagcaaag acagcaggaa gaggcaagtg
      181 gagacagtcc atttgggtgg ggaaatgact gggttgtcaa tggatagata gatagataga
      241 cagatagata gatagataga tagatagata gatagataga tagatagaca gacagacaga
      301 cagacagaca gatgaattta tttaacttcc cacttatttt tacatctctt attattcccg
      361 aaagactcca tggaggctta caaatacaga ccaaatgcaa taagataaca caatcctgta
      421 taataaaggg agcacccttg tcttgtgtgg agatggcagt agctttggta gcacatcttg
      481 gaggatctgg gctttctacc tccagtttgc cattaactgt gtgtttcagg caagtccctt
      541 caccttcctg ccctcgtttt ttctggtgta cccagtgccc tggccacgtc caggggctat
      601 tgtggaaagc aaatgagcag cctcaagttt ctttatctgt aaaacgccat tagtaggaat
      661 taagtgatgc agtgcattcc aagagcctgg cacgtggtcc atcttcacca aacgctggct
      721 ctgctgtgat gtttgcacga atatatattt cttggctcca gaagcagaag ggagctgaac
      781 gggtattgca gcgcctgatc tgcagggcca tgccattctc tccgtctcac tcattcgtgc
      841 ccattttcat tggagggtgg ctggcttttc catgtccaac agcaaacatt tgccaaaggc
      901 taatgcttgt catttcttca atcaacacac atataacagt aaaaatgatg ccactcaccg
      961 tgtgccaggc cctgggatat gctctttgca tgcattatct cacttcccct caccaccacc
     1021 ctaggagctg ggtactgcgc ccattttaca gatgaggaga ctgaggcaca gagaggtgaa
     1081 gtcattgact gagaccacac cattagcaag tgagagggcc agggtgtgaa ctcaagtgcc
     1141 aagccctgcc cctgcttggg gaggattcat ggatacagtg gctcccttca gaagggctca
     1201 agctcccctg ctgtccgctg gccactgacc ttggctgttt tgtggccatt gggcttcagc
     1261 cttttctgga ggccaattca atggactaga actcaacgag gattctacag tacctgtgac
     1321 tacccggcac aggagtcctg gggcgaagga ggtcacagcg gagtggggag gctgcctgcg
     1381 aggtgtggaa agcatctgcc tcctgcagca ggaggcctga ttgtcagtct ccacttcccc
     1441 cacactggct gtgtgaccct gggcaaggcc cctacctctc tgtgccttag acacctgcct
     1501 catcagtaaa atgggaaaca taaagcttga ggagaaaaaa cagtgaggaa agtgcttggt
     1561 aaaccataaa cgcagggggt tttccccacg atgaaagtct tgccctgtcc ttaaggatcc
     1621 tgcatgctcc atggtaaact ggcaaggaca caatccgaga gccaggccaa atggtggtag
     1681 gaggaagtgc cagggcggca gggaaggaga aattagttgg ggatgactga agcccttctg
     1741 aatgaggtgt aacctgcagg gtgcacgttg atgggtgaat aggatgacag atgaagggcc
     1801 cttctggagg atctactgct ggtcagcttt ggcggtgctc cagggatgtg gtgggggagc
     1861 agtgggccag gctcctaggg accactgcag gaaagcctcc agtaccacat aaggaacacc
     1921 agctgcagcg gcggcatcca tggtggctgg tgcggtgggc cctgcctggg ctgggggcca
     1981 ctgtgggcca ctgcccccag aaactcccaa gtacagtgta cactgaagag attgtgcaac
     2041 ttccactttc ctgttcccac cgccccagct gtagcttccc cagctggcag aacagcacga
     2101 ctgctgaccc atcccaaccc ttctctttgg aaatttcctt tgggaaggtt tggaaactca
     2161 gccagctcct ccctgcccct gcctccccac gcacgtcctc accctgcaga aggagcccct
     2221 ccatccacac tgcagctttt cttgtgacca gggtaaatat tacctggcac tgttaatgtt
     2281 taatgcccgc acctcgtccc cggccacccc gtggaggcag ccagacactc tccaaaaagc
     2341 agagacagca ggaagagggg agtggaggca gcccattcac ctggggaaga tctgcgatct
     2401 aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
     2461 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
     2521 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     2581 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
     2641 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     2701 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     2761 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
     2821 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
     2881 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     2941 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     3001 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     3061 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     3121 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     3181 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     3241 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     3301 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     3361 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     3421 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     3481 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     3541 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     3601 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     3661 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     3721 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     3781 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     3841 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     3901 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     3961 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     4021 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     4081 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     4141 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4201 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4261 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     4321 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     4381 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     4441 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     4501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6841 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     6901 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     6961 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     7021 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     7081 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     7141 tgcaggtgcc agaacatttc tctatcgata
//
