LOCUS       sequence-296327-        5162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXL1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5162)
  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 5162)
  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-FOXL1
FEATURES             Location/Qualifiers
     source          1..5162
                     /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    1214..1348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1238..1257)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1292..1311
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1461..1481)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1461..1479)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1500..1516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1500..1516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1513..1535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1524..1540
                     /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(1548..1554,1555..1572,1573..1578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1593..1614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1731..1748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1902..2490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1982..2001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2661..3452,3453..3521))
                     /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     3284..3303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3522..3626)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3652..4107)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3789..3810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4001..4020
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4178..5162
                     /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 tcgaattcag tcacctcttc gatccccggc tgcctgccct ggccgcctcg
      181 cccatgctgt atctgtacgg tcccgagaga cccggcctcc ctctggcctt cgcccccgcg
      241 gctgctctag ctgcctcggg ccgggccgag accccgcaga agcctcccta cagctacatc
      301 gcgctcatcg ccatggcgat ccaggacgcg cccgagcaga gggtcacgct caacggcatc
      361 taccagttca tcatggaccg cttccccttc taccacgaca accggcaggg ctggcagaac
      421 agcatccgcc acaacctctc gctcaacgac tgcttcgtca aggtgccccg cgagaaaggg
      481 cggccgggca agggcagcta ctggacgctg gacccccgct gcctggacat gtttgagaac
      541 ggcaactacc ggcgccggaa gaggaagccc aagccgggcc ccggggcccc ggaggccaag
      601 aggccccgcg ccgagacgca ccagcgcagc gcggaggcgc agccggaggc ggggagcggg
      661 gcagggggct cgggccccgc aatctcccgc ctgcaggcag cgcccgcggg cccctcgccc
      721 ctcctggacg gcccctctcc gccggcgccc ctccactggc cggggaccgc gtccccgaac
      781 gaggacgctg gtgacgctgc ccagggcgca gcggccgtgg cggtcgccca ggcagcgcgc
      841 acaggggacg gcccggggtc ctctctgcgc cccgcctccc gcagctctcc gaagagctcc
      901 gacaagtcca agagcttcag catagacagc atcctggcgg gaaagcaggg ccagaagccg
      961 ccttcagggg acgaactcct agggggtgcc aagcctgggc ccggcggccg tctgggtgcc
     1021 tcgctcctgg ccgcctcctc cagcctccgt ccgcctttca acgcttccct gatgctcgac
     1081 ccgcatgtcc agggcggctt ttaccagctc gggatcccct tcctctctta tttccccctg
     1141 caggttcccg acacggtact ccacttccag taaagcctcg agcctctaga actatagtga
     1201 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
     1261 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1321 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1381 ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc
     1441 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     1501 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1561 gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1621 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2341 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3661 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3721 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3781 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3841 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3901 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3961 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4021 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4081 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     4141 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
     4201 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
     4261 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
     4321 ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg
     4381 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata
     4441 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat
     4501 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta
     4561 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct
     4621 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc
     4681 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc
     4741 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct
     4801 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc
     4861 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg
     4921 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca
     4981 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
     5041 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
     5101 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
     5161 ac
//
