LOCUS       sequence-362709-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG tagged HNF1B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  AUTHORS   Beucher A, Miguel-Escalada I, Balboa D, De Vas MG, Maestro MA, 
            Garcia-Hurtado J, Bernal A, Gonzalez-Franco R, Vargiu P, Heyn H, 
            Ravassard P, Ortega S, Ferrer J
  TITLE     The HASTER lncRNA promoter is a cis-acting transcriptional 
            stabilizer of HNF1A.
  JOURNAL   Nat Cell Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8.
  PUBMED    36202974
REFERENCE   2  (bases 1 to 6851)
  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: "Nat Cell 
            Biol. 2022 Oct 6. pii: 10.1038/s41556-022-00996-8. doi: 
            10.1038/s41556-022-00996-8."
COMMENT     Sequence Label: pcDNA3.1-HNF1B-WT-Flag-UTR
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..31
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     3'UTR           35..160
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(226..243)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    232..456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      502..930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(589..608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     799..820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(939..959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        944..1273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1124..1259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1186..1205
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1321..1368
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1387..1761
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1891..2012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1928..1947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1982..2001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2061..2077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2061..2077)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2074..2096)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2085..2101
                     /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        complement(join(2109..2115,2116..2133,2134..2139))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2154..2175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2292..2309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2463..3051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2543..2562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3222..4013,4014..4082))
                     /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     3845..3864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4083..4187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4262..4281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4453..4832
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4833..5036
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4987..5007
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5081..5100
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5081..5099
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactagtgac tacaaagacg atgacgataa ataaaaccag cctcaagaac acccgaatgg
       61 agtctctaag ctacataata ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag
      121 ccattcgtat ctgctcctaa taaaaagaaa gtttcttcac ctgcagatat ccagcacagt
      181 ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
      241 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      301 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      361 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
      421 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
      481 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      781 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      841 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      901 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
      961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1201 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1261 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     1321 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     1381 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     1441 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     1501 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     1561 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     1621 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     1681 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     1741 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     1801 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     1861 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     1921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1981 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     2041 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2101 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2161 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2221 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4201 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     4261 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     4321 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     4381 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     4441 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     4501 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4561 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagyaa
     4621 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4681 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4741 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4801 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     4861 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4921 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     4981 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     5041 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     5101 agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat ccactagtcc
     5161 agtgtggtgg gcaccatggt gtccaagctc acgtcgctcc agcaagaact cctgagcgcc
     5221 ctgctgagct ccggggtcac caaggaggtg ctggttcagg ccttggagga gttgctgcca
     5281 tccccgaact tcggggtgaa gctggagacg ctgcccctgt cccctggcag cggggccgag
     5341 cccgacacca agccggtctt ccatactctc accaacggcc acgccaaggg ccgcttgtcc
     5401 ggcgacgagg gctccgagga cggcgacgac tatgacacac ctcccatcct caaggagctg
     5461 caggcgctca acaccgagga ggcggcggag cagcgggcgg aggtggaccg gatgctcagt
     5521 gaggaccctt ggagggctgc taaaatgatc aagggttaca tgcagcaaca caacatcccc
     5581 cagagggagg tggtcgatgt caccggcctg aaccagtcgc acctctccca gcatctcaac
     5641 aagggcaccc ctatgaagac ccagaagcgt gccgctctgt acacctggta cgtcagaaag
     5701 caacgagaga tcctccgaca attcaaccag acagtccaga gttctggaaa tatgacagac
     5761 aaaagcagtc aggatcagct gctgtttctc tttccagagt tcagtcaaca gagccatggg
     5821 cctgggcagt ccgatgatgc ctgctctgag cccaccaaca agaagatgcg ccgcaaccgg
     5881 ttcaaatggg ggcccgcgtc ccagcaaatc ttgtaccagg cctacgatcg gcaaaagaac
     5941 cccagcaagg aagagagaga ggccttagtg gaggaatgca acagggcaga atgtttgcag
     6001 cgaggggtgt ccccctccaa agcccacggc ctgggctcca acttggtcac tgaggtccgt
     6061 gtctacaact ggtttgcaaa ccgcaggaag gaggaggcat tccggcaaaa gctggccatg
     6121 gacgcctata gctccaacca gactcacagc ctgaaccctc tgctctccca cggctccccc
     6181 caccaccagc ccagctcctc tcctccaaac aagctgtcag gagtgcgcta cagccagcag
     6241 ggaaacaatg agatcacttc ctcctcaaca atcagtcacc atggcaacag cgccatggtg
     6301 accagccagt cggttttaca gcaagtctcc ccagccagcc tggacccagg ccacaatctc
     6361 ctctcacctg atggtaaaat gatctcagtc tcaggaggag gtttgccccc agtcagcacc
     6421 ttgacgaata tccacagcct ctcccaccat aatccccagc aatctcaaaa cctcatcatg
     6481 acacccctct ctggagtcat ggcaattgca caaagcctca acacctccca agcacagagt
     6541 gtccctgtca tcaacagtgt ggccggcagc ctggcagccc tgcagcccgt ccagttctcc
     6601 cagcagctgc acagccctca ccagcagccc ctcatgcagc agagcccagg cagccacatg
     6661 gcccagcagc ccttcatggc agctgtgact cagctgcaga actcacacat gtacgcacac
     6721 aagcaggaac ccccccagta ttcccacacc tcccggtttc catctgcaat ggtggtcaca
     6781 gataccagca gcatcagtac actcaccaac atgtcttcaa gtaaacagtg tcctctacaa
     6841 gcctgggaat t
//
