LOCUS       sequence-298987-        5517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXA2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5517)
  AUTHORS   Moparthi L, Koch S
  TITLE     A uniform expression library for the exploration of FOX 
            transcription factor biology.
  JOURNAL   Differentiation. 2020 Sep - Oct;115:30-36. doi: 
            10.1016/j.diff.2020.08.002. Epub 2020 Aug 20.
  PUBMED    32858261
REFERENCE   2  (bases 1 to 5517)
  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: 
            "Differentiation. 2020 Sep - Oct"; volume: "115"; pages: "30-36. 
            doi: 10.1016/j.diff.2020.08.002. Epub 2020 Aug 20"
COMMENT     Sequence Label: Flag-FOXA2
FEATURES             Location/Qualifiers
     source          1..5517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..117
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      298..307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1569..1703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1593..1612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1647..1666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1816..1836)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1816..1834)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1855..1871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1855..1871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1868..1890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1879..1895
                     /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(1903..1909,1910..1927,1928..1933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1948..1969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2086..2103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2257..2845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2337..2356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3016..3807,3808..3876))
                     /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     3639..3658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3877..3981)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4007..4462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4144..4165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4356..4375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4533..5517
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcagccacc atggattaca aggacgacga cgataaggga
      121 tcccatcgat tcgaattcca ctcggcttcc agtatgctgg gagcggtgaa gatggaaggg
      181 cacgagccgt ccgactggag cagctactat gcagagcccg agggctactc ctccgtgagc
      241 aacatgaacg ccggcctggg gatgaacggc atgaacacgt acatgagcat gtcggcggcc
      301 gccatgggca gcggctcggg caacatgagc gcgggctcca tgaacatgtc gtcgtacgtg
      361 ggcgctggca tgagcccgtc cctggcgggg atgtcccccg gcgcgggcgc catggcgggc
      421 atgggcggct cggccggggc ggccggcgtg gcgggcatgg ggccgcactt gagtcccagc
      481 ctgagcccgc tcggggggca ggcggccggg gccatgggcg gcctggcccc ctacgccaac
      541 atgaactcca tgagccccat gtacgggcag gcgggcctga gccgcgcccg cgaccccaag
      601 acctacaggc gcagctacac gcacgcaaag ccgccctact cgtacatctc gctcatcacc
      661 atggccatcc agcagagccc caacaagatg ctgacgctga gcgagatcta ccagtggatc
      721 atggacctct tccccttcta ccggcagaac cagcagcgct ggcagaactc catccgccac
      781 tcgctctcct tcaacgactg tttcctgaag gtgccccgct cgcccgacaa gcccggcaag
      841 ggctccttct ggaccctgca ccctgactcg ggcaacatgt tcgagaacgg ctgctacctg
      901 cgccgccaga agcgcttcaa gtgcgagaag cagctggcgc tgaaggaggc cgcaggcgcc
      961 gccggcagcg gcaagaaggc ggccgccgga gcccaggcct cacaggctca actcggggag
     1021 gccgccgggc cggcctccga gactccggcg ggcaccgagt cgcctcactc gagcgcctcc
     1081 ccgtgccagg agcacaagcg agggggcctg ggagagctga aggggacgcc ggctgcggcg
     1141 ctgagccccc cagagccggc gccctctccc gggcagcagc agcaggccgc ggcccacctg
     1201 ctgggcccgc cccaccaccc gggcctgccg cctgaggccc acctgaagcc ggaacaccac
     1261 tacgccttca accacccgtt ctccatcaac aacctcatgt cctcggagca gcagcaccac
     1321 cacagccacc accaccacca gccccacaaa atggacctca aggcctacga acaggtgatg
     1381 cactaccccg gctacggttc ccccatgcct ggcagcttgg ccatgggccc ggtcacgaac
     1441 aaaacgggcc tggacgcctc gcccctggcc gcagatacct cctactacca gggggtgtac
     1501 tcccggccca ttatgaactc ctcttaagaa gacgacggct ctagaactat agtgagtcgt
     1561 attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1621 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1681 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1741 agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt
     1801 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     1861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag
     1921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2041 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2101 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2161 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2221 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2281 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2341 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2401 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2461 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2521 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2581 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2641 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2701 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2761 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2821 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2881 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2941 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3001 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3061 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     4021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4441 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4501 cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata
     4561 tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata
     4621 tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact
     4681 tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac
     4741 ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa
     4801 ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca
     4861 atatccaata tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc
     4921 cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa
     4981 tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc actcccccat
     5041 tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg
     5101 acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg
     5161 cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt
     5221 catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta
     5281 catcaatatc tattaatagt aacttggcaa gtacattact attggaagta cgccagggta
     5341 cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta
     5401 catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa
     5461 atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
