LOCUS       sequence-299374-        5384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXI3in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5384)
  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 5384)
  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-FOXI3
FEATURES             Location/Qualifiers
     source          1..5384
                     /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    1436..1570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1460..1479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1514..1533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1683..1703)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1683..1701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1722..1738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1722..1738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1735..1757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1746..1762
                     /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(1770..1776,1777..1794,1795..1800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1815..1836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1953..1970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2124..2712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2204..2223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2883..3674,3675..3743))
                     /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     3506..3525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3744..3848)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3874..4329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4011..4032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4223..4242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4400..5384
                     /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 tcgaattcgc cctctactgc ggcgacaact tcggagtgta ttcgcagccc
      181 ggcctgcccc cgcccgccgc caccgccgcc gccccgggcg cccccccggc agccagggcg
      241 ccttacgggc tggccgacta cgccgcgccg ccggccgccg ccgccaaccc ctacctgtgg
      301 ctcaacgggc ccggcgtggg aggcccgccc tccgccgccg ccgcagccgc cgccgcgtac
      361 ctgggcgctc cgccgccgcc gccgcccccc ggggccgcgg ccgggccctt tctgcagcca
      421 ccgcctgccg ccggcacctt cggctgctct cagcggccct tcgcgcagcc cgcgcccgcc
      481 gcgcccgcct cgcccgccgc gcccgcgggg cccggggagc tgggctggct gtccatggcc
      541 agccgcgagg acctgatgaa gatggtgcgg ccgccctact cgtattcggc gctcatcgcc
      601 atggccattc agagcgcgcc cgagcgcaaa ctcactctca gccacatcta ccagttcgtc
      661 gccgatagct tccccttcta ccagcgcagc aaggccggct ggcagaactc catccgccac
      721 aacctgtcgc tcaacgactg cttcaagaag gtgccccgcg acgaggacga cccaggaaag
      781 ggtaattatt ggactcttga tccgaactgc gagaaaatgt ttgacaatgg gaacttccgt
      841 cggaagcgaa agcgccgctc tgaggccagc aatggctcca cagtggctgc tgggacatca
      901 aagtccgaag aagggctctc ctcaggattg gggtctggag tgggtgggaa gccagaagaa
      961 gagagcccct ccactctgct gaggccttcc cactccccag agcctccgga gggcaccaag
     1021 agtactgcct catctcctgg aggacccatg ctcacctcca ccccttgtct caacactttc
     1081 ttcagcagcc tcagctcctt gagtgtcagc agcagtgtga gtacccagcg ggctctccct
     1141 ggcagccgcc acctagggat ccagggggcc cagctgccct ccagcggcgt gttctccccg
     1201 acctccatct cagaggcctc agcagacacc ttgcaactga gcaatagcac cagcaatagc
     1261 accggccaga gatcttccta ttacagccct ttccctgcca gcaccagcgg ggggcaaagc
     1321 agccccttca gcagcccttt ccacaacttt agcatggtga acagcctcat ctacccccga
     1381 gagggctccg aggtgtagct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     1441 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1501 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1561 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1621 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     1681 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     1741 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     1801 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1861 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1921 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1981 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2041 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2101 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2161 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2221 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2281 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2521 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2821 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2881 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2941 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3001 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3061 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3121 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3181 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3241 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3301 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3361 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3421 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3481 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3541 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3601 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3661 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3721 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3781 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3841 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     3901 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3961 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4021 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4081 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4141 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4201 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4261 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4321 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4381 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     4441 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     4501 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     4561 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     4621 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     4681 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     4741 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     4801 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     4861 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     4921 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     4981 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     5041 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     5101 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     5161 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     5221 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     5281 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     5341 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
