LOCUS       sequence-120281-        8299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8299)
  AUTHORS   Shin M, Male I, Beane TJ, Villefranc JA, Kok FO, Zhu LJ, Lawson ND
  TITLE     Vegfc acts through ERK to induce sprouting and differentiation of 
            trunk lymphatic progenitors.
  JOURNAL   Development. 2016 Oct 15;143(20):3785-3795. Epub 2016 Sep 12.
  PUBMED    27621059
REFERENCE   2  (bases 1 to 8299)
  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: 
            "Development."; date: "2016-10-15"; volume: "143(20)"; pages: 
            "3785-3795. Epub 2016 Sep 12"
COMMENT     Sequence Label: pCS-flt4um159-HA
FEATURES             Location/Qualifiers
     source          1..8299
                     /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"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4231..4255)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4257..4283
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4288..4308)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    4348..4482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4372..4391)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4426..4445
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4595..4615)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4595..4613)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4634..4650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4634..4650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4647..4669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4658..4674
                     /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(4682..4688,4689..4706,4707..4712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4727..4748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4865..4882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5036..5624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5116..5135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5795..6586,6587..6655))
                     /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     6418..6437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6656..6760)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6786..7241)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6923..6944)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7135..7154
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7315..8299
                     /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 atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctccgaattc gcccttgcca ccatgaagag agattttacg tttttctgtc
      181 ggatttggat tgggattccc ttcttctcag gtctggtgaa tgggttttct atgagtccac
      241 ccacccttga caacaccaaa gaccagcttg tgattaacgc taacgacaca ctgaacatta
      301 catgcagggg tcagcgaatt ttggactgga gctggcctga agagtctttg agtaaagtgg
      361 agtttactga tcgccagggt cagcagtcac ccactgacac cccagggtat agagaaatca
      421 ggctgaagga gtgtcaaggg gtggctggaa aaccctactg caagatctta atacttacca
      481 atgcccaggc caatgactcg ggttattacc gctgcttcta caaggacatt aaagctgtta
      541 ttgatggcac cacggcagcc agcatttttg tgtttgttcg agacccagag catccattca
      601 tcaaaagggg agacaacgac atggagacca tcttcataac agactctgaa acacacattg
      661 aagtaccatg tctggtttca gatcccgacc taaaagtcac tctcttttcg ttagtgccgt
      721 atccggagcc tgtggatggc agtgtggtca cctggaataa taaaaagggt tggtcgattc
      781 ccaggcatat cattcagaac acttccacct ttattggctt ctactgctcc atctcagtcc
      841 aaaacagcca gcacacatca tcaatctatg ttgtccaggt cattgggctg aagttttatg
      901 agttcaagct gtttcctgaa gactcgcctg tggagttgat gcagggggag agtttggtgc
      961 tcaactgcac tgcactggtc gacttcaaca ctggtgtgga tttccagtgg gactatccag
     1021 gcaaaaagga gaaccggttg gcgagtttac agcctctgcg taacgtcctg gatgaagcca
     1081 cggaaatctc cagtatcctc tcaatcagaa acattcactt ggatgatagt ggctactaca
     1141 cctgctgggc taatactctg gaaatgaaac gagagctcac tactgtagtc atagttcatg
     1201 aaaaaccttt tatcagcttg gactacagga atggatcagt tatagaggca aaagagggac
     1261 aaaaatctgt ccggctatcc gtaaaagtgt cagcataccc ttcaccagaa atacaatggt
     1321 ataagaatgg aaaactgatc agcagcaaga attcaagcag gtttaaggtc cagcagcact
     1381 ctctacagat cagggatgtg tgtaagcagg atgctggaga atacatgctg gtcctgaaaa
     1441 atagccctgc agccctggag aaaagactca actttactct catagtcaac gttccacctc
     1501 agattcatga gaaggaggca gcaccaccta caaacctcta tgggaagggc actcgacaga
     1561 tcctcacttg caccgcagac gggagccctc cagcctccat cagctggcag tggaggccct
     1621 ggagcccctg cgatctggag cgtacccgca gagccctgag acggaggggt ggacgagacc
     1681 agagtccttt ctgtcacaac tggatggatt tggaccctga acatgctgta aaccctattg
     1741 agagtattga cacattgact cagatggtgg acggcaaaga aaagactgtc ggccgtgtgg
     1801 ttattcagaa tgccagtgtg ccagctatgt tggctgagaa caaagtggga aaagatgaac
     1861 gactgattta tttctacgta accactatcc ctgaaggatt cgatatagag atggagccct
     1921 cagaggatcc gcttgagcag gacctggtac agctgaagtg taatgcagat aatttcacct
     1981 atgagaacct gcggtggtac cgcctggacc cgcagactgt tcctccagag ctggactgca
     2041 agagtctgca ccagtatgcc acatttttgg agggacaact atcttttcag accaccagca
     2101 acaactgggt cctgcagctt aacattacca acattcagct acaggatgag gggaactatg
     2161 tgtgtgaggt gcagaaccga cgaaccgggg tgaaacattg tcaccgcaaa tacatcccag
     2221 tcaaagcaat ggaagcacct cgataccgtc acaacccaac taatcatacg gtaaatgtga
     2281 gcgagtcgct gcagatgaac tgtgatgtgg aaggcacacc ctttcctcag ctgtcctggt
     2341 ttaaagataa ccaacccctc catcagatat cagggatcct ccttcaggac tccaatcgga
     2401 ctctcagcat tcagcgagtg cgagaggaag acgccggtct gtacacctgc tctgcctgca
     2461 accagaaggg ctgcgttcag tcctccgcca cggtgtcagt gattggctct gatgacaaaa
     2521 ccaatgtaga aatagtgatc ctcatcggca caggagtcat cgccatcttt ttctgggtcc
     2581 ttcttcttgt catcttctgc aatgtcaagc gagtgaaccc agctgatatc aagacaggct
     2641 atctgtccat catcatggac ccgggggaag tgccgctgga ggagcagtgt gaatacctgc
     2701 cgtacgactc cagccagtgg gagatttcca gagacagact gcgcctcggt aaggtgctcg
     2761 gccatggagc tttcgggaag gtgattgaag cctcaatctt tggtcatgac aagaaatcct
     2821 ctgcaaacac agtggctgtc aagatgttga aagagggagc aacagccagc gagcacaaag
     2881 ccctgatgtc agagctgaag atcctcattc atattggaaa ccatctcaat gttgtcaacc
     2941 ttcttggagc ttgtaccaaa cctaacggtc ctctcatggt tatagtggag tattgtaaat
     3001 atggaaatct gtccaacttc ctgcgtgcca agagggagtt ttttttacca tacagggatc
     3061 gttctcctaa aacccaaagt caagtgcgac ggatgataga agcaggtcag gcgagccaga
     3121 gtgagcatca gccgtccact tcctccacca accctccacg tgttacagtg gatgatctct
     3181 ggaaaactcc actcacaata gaggatctga tatgctacag ttttcaagtt gcacgaggaa
     3241 tggagtttct ggcatctcgt aagtgtattc atcgagatct ggcagcccga aacatcctcc
     3301 tgtccgagaa caatgtggtg aagatctgtg actttggttt agcacgggac atctacaaag
     3361 accctgacta tgtgcgcaaa ggcaatgcca ggctgccgct gaagtggatg gctccagaga
     3421 gcatctttga taaggtttac accagtcaga gtgacgtctg gtcttttgga gttctgctct
     3481 gggagatttt ctcactagga gcgtctccat accccggcat ccagattgat gaagatttct
     3541 gcaaacgact caaagacggt accaggatga gagcaccaga caatgcgtcc cctgaaatat
     3601 atggcatcat gttggcctgc tggcagggcg agcccaggga gagacccaca tttccagctc
     3661 tggtggagat acttggagat ctactgcagg aaaacagtct accagaaatc ccgttcaatg
     3721 tgtctcagag ctcagaggac gatggcttct cacaagcttc ttccagacca ccatcacaag
     3781 aggagatcag attggcctgc aacacactgc caacacgata ttacaactgc gtgccatttg
     3841 ctggctgtgt gatggtcggg ccctccagca catgtcattc tcgagtgaag acctttgagg
     3901 aacttcctat ggaaatgaca tcacacaaga cacagcatga cagtcagaca gacagtggga
     3961 tggttttggc atcagatgaa ctggagaggt ttgaacacaa gcacagagga gccatgttga
     4021 cacatgcgac gacgggccaa agcactgagc gcctcatcag ttgtccatcg gtgagcagca
     4081 gcggcagcgg tggtggtttg ctccgtccag tgttcttcac tcaactctca ggccagacct
     4141 tctacaacaa cgagtacggt catctgtcag aggagggcgt cagcgactac ttctcctcct
     4201 cagaccaggc cgtttacaag ggcgaattcg acccagcttt cttgtacaaa gtggactatc
     4261 cgtacgacgt accagactac gcataaccaa ctttattata catagttgca ctcgagcctc
     4321 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     4381 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4441 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4501 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     4561 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     4621 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4681 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4741 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4861 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6781 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     6841 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     6901 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     6961 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7021 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7081 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7141 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7201 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     7261 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat
     7321 agccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     7381 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     7441 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg
     7501 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     7561 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     7621 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     7681 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     7741 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     7801 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     7861 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     7921 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     7981 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     8041 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     8101 ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     8161 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     8221 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     8281 caatgctcgt ttagggaac
//
