LOCUS       sequence-426199-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human ALX4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  AUTHORS   Kim S, Morgunova E, Naqvi S, Goovaerts S, Bader M, Koska M, Popov A,
            Luong C, Pogson A, Swigut T, Claes P, Taipale J, Wysocka J
  TITLE     DNA-guided transcription factor cooperativity shapes face and limb 
            mesenchyme.
  JOURNAL   Cell. 2024 Feb 1;187(3):692-711.e26. doi: 
            10.1016/j.cell.2023.12.032. Epub 2024 Jan 22.
  PUBMED    38262408
REFERENCE   2  (bases 1 to 6099)
  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"; doi: 
            "10.1016/j.cell.2023.12.032"; journalName: "Cell"; date: "2024-02-1-
            1"; volume: "187"; issue: "3"; pages: "692-711.e26"
COMMENT     Sequence Label: pCAG_ALX4-Flag-HA
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    878..899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(914..919,920..937,938..944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    952..968
                     /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)."
     primer_bind     957..979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     976..992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     976..992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1012..1391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1393..1667
                     /label=chicken beta-actin promoter
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2697..2716
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2697..2715
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3963..3986
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3987..4013
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4024..4041)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4030..4254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4326..4343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4326..4342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4335..4357)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4483..4938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4620..4641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4832..4851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4965..5069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5070..5138,5139..5930)
                     /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     complement(5288..5307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      901 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      961 gataacaatt tcacacagga aacagctatg accatgaggc gcgccggatt cgacattgat
     1021 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     1081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     1141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     1201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1381 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     1441 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     1501 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     1561 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagatcctt aattaataat
     2701 acgactcact atagggggtc gacatgaatg ctgagacttg cgtctcttac tgcgagtcgc
     2761 cggccgctgc catggacgcc tactacagcc cggtgtcgca gagtcgggag ggctcgtcgc
     2821 cttttagggc atttcccgga ggcgacaagt tcggcacaac tttcctgtcg gccgccgcca
     2881 aagcacaggg attcggggac gccaagagcc gggcccgtta cggcgctggg cagcaggacc
     2941 tggcgacacc cctggagagt ggagctgggg cgcggggctc ctttaacaag ttccagcccc
     3001 agccgtcgac cccgcagccc cagccgccgc cgcagccgca gccgcagcag cagcagccgc
     3061 agccccagcc gcctgcgcaa ccgcatcttt acttgcagcg aggcgcctgc aagacgcccc
     3121 cggacggcag cctcaaactc caggaaggca gcagcggcca cagcgcggcc ttgcaggttc
     3181 cctgctacgc taaagagagc tccctgggtg agccagagtt accccctgac tctgacactg
     3241 tggggatgga cagcagctac ctgagtgtca aggaggctgg ggtgaagggg ccccaggacc
     3301 gggccagctc agacctcccc agcccattgg agaaggccga ctcggagagc aacaagggca
     3361 agaagcggcg gaaccggacc accttcacca gctaccagct ggaggagctg gagaaggtct
     3421 tccagaagac ccactaccca gacgtgtatg cgcgggaaca gctggccatg aggacagacc
     3481 tcactgaggc ccgcgtgcag gtctggttcc agaaccgaag ggccaagtgg aggaagcggg
     3541 agcgttttgg gcagatgcag caggttcgaa cccacttctc cactgcatat gagctgcccc
     3601 tcctcacccg agctgagaac tacgcccaga ttcagaaccc gtcctggctc ggcaacaacg
     3661 gggctgcctc accagtgcca gcctgcgtgg tcccctgcga cccggtgcct gcctgcatgt
     3721 cccctcatgc ccacccccct ggctctgggg ccagcagcgt caccgacttc ctgagtgtgt
     3781 ctggggctgg cagtcatgtg ggccagacgc acatgggcag cctgtttgga gcagccagcc
     3841 tcagcccagg cctcaatggc tacgagctca acggcgagcc ggaccgcaag acctcgagca
     3901 tcgcggccct ccgcatgaag gccaaggagc acagtgcggc catttcctgg gccacatgta
     3961 cagactacaa agacgacgac gacaaatacc catacgacgt gccagactac gcctgatgat
     4021 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4081 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4141 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4201 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4261 aggcggaaag aaccagctgg ggctcgagat ccactagttc tagcctcgag gctagagcgg
     4321 ccgccactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4381 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4441 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata
     4501 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4561 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4621 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4681 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4741 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4801 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4861 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4921 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5041 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5161 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5221 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5281 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5341 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5461 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5521 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5581 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5641 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5701 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5761 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5821 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5881 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5941 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6001 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6061 actgagcgtc agaccccgta gaaaagatca aaggatctt
//
