LOCUS       sequence-462715-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A middle entry gateway clone containing V5 tagged human FOXF1 and an
            H2A tagged mCherry, separated by a P2A peptide cleavage sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  AUTHORS   Cantu Gutierrez ME, Hill MC, Largoza G, Martin JF, Wythe JD
  TITLE     Defining the Transcriptional and Epigenetic Basis of Organotypic 
            Endothelial Diversity in the Developing and Adult Mouse
  JOURNAL   bioRxiv 2021.11.15.468651
REFERENCE   2  (bases 1 to 4928)
  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: "bioRxiv 
            2021.11.15.468651"
COMMENT     Sequence Label: JDW 1391 (pME-V5-FOXF1-P2A-H2A-mCherry)
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      675..684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             684..707
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1872..1928
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2325..3035
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2470..2488)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2738..2757
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2932..2955
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(3053..3152)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3169..3188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3170..3188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3193..3209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3193..3209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3311..3330)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3322..4131
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3389..3408)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4278..4866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4767..4786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg atccgccacc atggattaca aggacgacga cgataaggga tcccatcgat
      721 tcgaattctc ttcggcgccc gagaagcagc agccaccgca cggcggcggc ggcggcggcg
      781 gcgggggagg cggcgcggcc atggaccccg cgtcgtccgg cccgtccaag gccaagaaga
      841 ccaacgccgg catccggcgc ccggagaagc cgccctattc ctacatcgcg ctcatcgtca
      901 tggccatcca gagttcaccc accaagcgcc tgacgctgag cgagatctac cagttcctgc
      961 agagccgctt ccccttcttc cggggctcct accagggctg gaagaactcc gtgcgccaca
     1021 acctctcgct caacgagtgc ttcatcaagc tacccaaggg ccttgggcgg cccggcaagg
     1081 gccactactg gaccatcgac ccggccagcg agttcatgtt cgaggagggc tcctttcggc
     1141 ggcggccgcg cggcttccga aggaaatgcc aggcgctcaa gcccatgtac agcatgatga
     1201 acgggctcgg cttcaaccac ctcccggaca cctacggctt ccagggctcg gccggcggcc
     1261 tctcgtgccc gcccaacagc ctggcgctgg agggcggcct gggcatgatg aacggccact
     1321 tgccgggcaa cgtggacggc atggccctgc ccagccactc ggtgccccac ctgccttcca
     1381 acggcggcca ctcgtacatg ggcggctgcg gcggcgcggc ggccggcgag tacccgcacc
     1441 acgacagctc ggtgcccgcc tccccgctgc tgcccaccgg cgccggtggg gtcatggagc
     1501 cgcacgccgt ctactcgggc tcggcggcgg cctggccgcc ctcggcgtcc gcggcgctca
     1561 acagcggcgc ctcttatatc aagcagcagc ccctgtcccc ctgtaacccc gcggccaacc
     1621 ccctgtccgg cagcctctcc acgcactccc tggagcagcc gtatctgcac cagaacagcc
     1681 acaacgcccc agccgagctg caaggcatcc cgcggtatca ctcgcagtcg cccagcatgt
     1741 gtgaccgaaa ggagtttgtc ttctctttca acgccatggc gtcctcttcc atgcactcgg
     1801 ccggcggggg ctcctactac caccagcagg tcacctacca agacatcaag ccttgcgtga
     1861 tgggtaccgg agccactaac ttctccctgt tgaaacaagc aggggatgtc gaagagaatc
     1921 ccgggccacc taggatggca ggtggaaaag caggtaaaga cagtggaaaa gccaagacga
     1981 aagccgtctc cagatctcag agagctggtc tgcagttccc tgtaggtcgt atccacaggc
     2041 acctgaagac acgaaccacc agccatggcc gtgtcggagc tacagctgcg gtttacagtg
     2101 ctgctatcct ggagtatctc accgctgagg tgctggagtt ggctggaaac gcttctaagg
     2161 atctgaaggt gaagcgaatc actcccagac atttacagct ggccatccgc ggagacgagg
     2221 agctggactc gcttatcaag gccaccattg ctggaggagg agtcattcct cacatccaca
     2281 agtctctgat tggtaagaag ggccagcaga aaaccgcagg ggggatggtg agcaagggcg
     2341 aggaggacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     2401 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2461 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     2521 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2581 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2641 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2701 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2761 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2821 tcaagcagag gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct
     2881 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagctggaca
     2941 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
     3001 actccaccgg cggcatggac gagctgtaca agtaatgata gctcgagaga tctcccagct
     3061 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     3121 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     3181 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     3241 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     3301 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     3361 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     3421 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     3481 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     3541 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3601 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     3661 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3721 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3781 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3841 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3901 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3961 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4021 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4081 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4141 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4201 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4261 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4321 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4381 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4801 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4861 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4921 cacatgtt
//
