LOCUS       sequence-14279-e        6086 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 6086)
  AUTHORS   Senkel S, Lucas B, Klein-Hitpass L, Ryffel GU
  TITLE     Identification of target genes of the transcription factor HNF1beta 
            and HNF1alpha in a human embryonic kidney cell line.
  JOURNAL   Biochim Biophys Acta. 2005 Dec 20. 1731(3):179-90.
  PUBMED    16297991
REFERENCE   2  (bases 1 to 6086)
  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: "Biochim 
            Biophys Acta. 2005 Dec 20. 1731(3):179-90."
COMMENT     Sequence Label: OPN(-213)-luc
FEATURES             Location/Qualifiers
     source          1..6086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1350..1359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1356..3008
                     /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(1422..1440)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3049..3170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3060..3079)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3114..3133
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3418..3435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3589..4177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3669..3688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4348..5139,5140..5208))
                     /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     4971..4990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5209..5313)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5340..5795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5427..5446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5637..5658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5926..5974
                     /note="synthetic polyadenylation signal"
     misc_feature    5988..6079
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6028..6047
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tctgctatcc ctgaattctg ctctaatatt cggactttcc ctgtaatttt
       61 ctttcattca gacacctttt aaatacctag taaagtgttt tttaatacag aaatttttaa
      121 aaatgttttt ctttttaagt ggcctacttt acataccttg ggagaaaaac tagaaaaaaa
      181 gatgattcca aaatcgaatc tgttccttta gaaatgtgca aaatttcctt attgatgcat
      241 acaatttaaa gatcttacgt ctactctcat tttaataacc tgttctttta aaggacatta
      301 caattcgtga ctgcctgccc ctcttaaaaa tttcataata gttaacacac atatagtcct
      361 taagatacgc agagcatttg catctaatat gtgctaagca ttgctagttt aacatactaa
      421 ttcatttaaa cccctcaaaa accccatgac ctaggtaata gtattgcatt tcatggatga
      481 gggaacaagg ataggtaggc tgggcgattt gcccaaggtt gcacaggtca gcagtgacac
      541 agcggaattc agaaccacgg tctggctcct gaagcagccc tctcaagcag tcatcctgct
      601 ctcagtcaga aactgcttta cttctgcaac atctagaata aattaccatt cttctatttc
      661 atatagaatt ttatatttta atgtcactag tgccatttgt ctaagtaaca agctactgca
      721 tactcgaaat cacaaagcta agcttgagta gtaaaggaca gaggcaagtt ttctgaactc
      781 cttgcaggct tgaacaatag ccttctggct cttcaataag tacaatcata caggcaagag
      841 tggttgcaga tattaccttt atgttactta aaccgaaaga aacaaaaatc cattctattt
      901 aattttacat taatgttttt ccctactttc tccctttttc atgggatccc taagtgctct
      961 tcctggatgc tgaatgccca tcccgtaaat gaaaaagcta gttaatgata ttgtacataa
     1021 gtaatgtttt aactgtagat tgtgtgtgtg cgtttttgtt tttttttgtt ttaaccacaa
     1081 aaccagaggg ggaagtgtgg gagcaggtgg gctgggcagt ggcagaaaac ctcatgacac
     1141 aatctctccg cctccctgtg ttggtggagg atgtctgcag cagcatttaa attctgggag
     1201 ggcttggttg tcagcagcag caggaggagg cagagcacag catcgtcggg accagactcg
     1261 tctcaggcca gttgcagcct tctcagccaa acgccgaccc tcgagatctg cgatctaagt
     1321 aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga
     1381 aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg
     1441 ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg
     1501 tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac
     1561 gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct
     1621 ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt
     1681 ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt
     1741 ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa
     1801 ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca
     1861 catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg
     1921 acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg
     1981 ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca
     2041 atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa
     2101 tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg
     2161 aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc
     2221 caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt
     2281 tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca
     2341 agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta
     2401 ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt
     2461 ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg
     2521 aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca
     2581 acgccttgat tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag
     2641 acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg
     2701 tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg
     2761 tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc
     2821 acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg
     2881 cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa
     2941 aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg
     3001 ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     3061 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3121 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3181 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     3241 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     3301 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3361 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     3421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3481 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3541 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3601 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3661 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3721 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3781 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3841 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3901 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3961 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4021 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4081 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4141 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4261 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4321 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4381 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4441 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4501 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4561 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4621 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4681 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4741 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4801 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4861 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4921 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4981 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5041 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5101 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5161 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5221 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5281 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5461 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5701 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5761 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     5821 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     5881 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
     5941 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
     6001 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     6061 ggtgccagaa catttctcta tcgata
//
