LOCUS       sequence-298986-        5545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXA1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5545)
  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 5545)
  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-FOXA1
FEATURES             Location/Qualifiers
     source          1..5545
                     /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"
     polyA_signal    1597..1731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1621..1640)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1675..1694
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1844..1864)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1844..1862)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1883..1899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1907..1923
                     /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(1931..1937,1938..1955,1956..1961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1976..1997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2114..2131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2285..2873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2365..2384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3044..3835,3836..3904))
                     /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     3667..3686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3905..4009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4035..4490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4384..4403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4561..5545
                     /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 tcgaattctt aggaactgtg aagatggaag ggcatgaaac cagcgactgg
      181 aacagctact acgcagacac gcaggaggcc tactcctccg tcccggtcag caacatgaac
      241 tcaggcctgg gctccatgaa ctccatgaac acctacatga ccatgaacac catgactacg
      301 agcggcaaca tgaccccggc gtccttcaac atgtcctatg ccaacccggg cctaggggcc
      361 ggcctgagtc ccggcgcagt aaccggcatg ccggggggct cggcgggcgc catgaacagc
      421 atgactgcgg ccggcgtgac ggccatgggt acggcgctga gcccgagcgg catgggcgcc
      481 atgggtgcgc agcaggcggc ctccatgaat ggcctgggcc cctacgcggc cgccatgaac
      541 ccgtgcatga gccccatggc gtacgcgccg tccaacctgg gccgcagccg cgcgggcggc
      601 ggcggcgacg ccaagacgtt caagcgcagc tacccgcacg ccaagccgcc ctactcgtac
      661 atctcgctca tcaccatggc catccagcag gcgcccagca agatgctcac gctgagcgag
      721 atctaccagt ggatcatgga cctcttcccc tattaccggc agaaccagca gcgctggcag
      781 aactccatcc gccactcgct gtccttcaat gactgcttcg tcaaggtggc acgctccccg
      841 gacaagccgg gcaagggctc ctactggacg ctgcacccgg actccggcaa catgttcgag
      901 aacggctgct acttgcgccg ccagaagcgc ttcaagtgcg agaagcagcc gggggccggc
      961 ggcgggggcg ggagcggaag cgggggcagc ggcgccaagg gcggccctga gagccgcaag
     1021 gacccctctg gcgcctctaa ccccagcgcc gactcgcccc tccatcgggg tgtgcacggg
     1081 aagaccggcc agctagaggg cgcgccggcc cccgggcccg ccgccagccc ccagactctg
     1141 gaccacagtg gggcgacggc gacagggggc gcctcggagt tgaagactcc agcctcctca
     1201 actgcgcccc ccataagctc cgggcccggg gcgctggcct ctgtgcccgc ctctcacccg
     1261 gcacacggct tggcacccca cgagtcccag ctgcacctga aaggggaccc ccactactcc
     1321 ttcaaccacc cgttctccat caacaacctc atgtcctcct cggagcagca gcataagctg
     1381 gacttcaagg catacgaaca ggcactgcaa tactcgcctt acggctctac gttgcccgcc
     1441 agcctgcctc taggcagcgc ctcggtgacc accaggagcc ccatcgagcc ctcagccctg
     1501 gagccggcgt actaccaagg tgtgtattcc agacccgtcc taaacacttc ctagctcccc
     1561 tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     1621 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1681 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1741 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     1801 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     1861 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1921 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     1981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4021 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4081 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4141 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4201 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4261 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4321 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4381 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4441 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     4501 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4561 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
     4621 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
     4681 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
     4741 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
     4801 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
     4861 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
     4921 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
     4981 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
     5041 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
     5101 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
     5161 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
     5221 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
     5281 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
     5341 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
     5401 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
     5461 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
     5521 tataagcaat gctcgtttag ggaac
//
