LOCUS       sequence-298996-        5101 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of Flag tagged human FOXB1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  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 5101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-FOXB1
FEATURES             Location/Qualifiers
     source          1..5101
                     /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    1153..1287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1177..1196)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1231..1250
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1400..1420)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1400..1418)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1439..1455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1439..1455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1452..1474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1463..1479
                     /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(1487..1493,1494..1511,1512..1517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1532..1553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1670..1687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1841..2429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1921..1940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2600..3391,3392..3460))
                     /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     3223..3242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3461..3565)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3591..4046)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3728..3749)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3940..3959
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4117..5101
                     /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 tcgaattccc tcggcccggc cgcaacacgt acagcgacca gaagccgccc
      181 tactcgtaca tctcgctgac cgctatggcc atccagagct ctcccgagaa gatgctgccg
      241 ctgagcgaga tctacaagtt catcatggac cgcttcccct actacaggga gaacacgcag
      301 cgctggcaga acagtctgcg ccacaacctc tccttcaacg actgcttcat caagatcccg
      361 cggcggccgg accagccagg caagggcagc ttctgggcgc tgcacccaag ctgcggggac
      421 atgttcgaga acggcagctt cctgcggcgc cgcaagcgct tcaaggtgct taagtccgac
      481 cacctggcgc ccagcaagcc agccgacgcg gcgcagtacc tgcagcagca ggccaagctg
      541 cggctcagcg cgctggcggc ctcgggcacg cacctgccac agatgcccgc cgccgcctac
      601 aacttgggcg gcgtggcgca gccctcgggc ttcaagcacc ccttcgccat cgagaacatc
      661 atcgcgcggg aatacaagat gcctgggggg ctggccttct ccgccatgca gccggtgccc
      721 gctgcctacc cgctccccaa ccagttgact accatgggca gctcgctggg caccggctgg
      781 ccacacgtgt atggctccgc cggcatgatc gactcggcca cccccatctc catggcgagt
      841 ggcgactaca gcgcctacgg cgtgccgttg aagccgctgt gccacgcggc gggccaaacg
      901 ctgcccgcca tccccgtgcc cattaagccc acgccggccg ccgtgcccgc gctgcctgcg
      961 ctgccagcgc ccatccccac cttgctctcg aactcgccgc cctcgctcag ccccacgtcc
     1021 tcgcaaacag ccaccagcca aagcagcccc gccaccccca gcgaaacgct caccagcccg
     1081 gcctccgcct tgcactcggt ggcggtgcac tgaccctcga gcctctagaa ctatagtgag
     1141 tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca aaccacaact
     1201 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1261 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1321 gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc
     1381 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     1441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1501 ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     3601 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3661 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3721 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3781 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3841 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3901 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3961 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4021 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     4081 agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg
     4141 tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg
     4201 tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg
     4261 gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt
     4321 ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat
     4381 tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata
     4441 ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat
     4501 tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc
     4561 ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc
     4621 ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct
     4681 attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc
     4741 aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc
     4801 cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc
     4861 agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag
     4921 ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca
     4981 agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac
     5041 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa
     5101 c
//
