LOCUS       sequence-332637-        4242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A vector with two different MCSs flanking an attP integration site 
            and a floxable pax-GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4242)
  AUTHORS   Yu JJS, Maugarny-Cales A, Pelletier S, Alexandre C, Bellaiche Y, 
            Vincent JP, McGough IJ
  TITLE     Frizzled-Dependent Planar Cell Polarity without Secreted Wnt 
            Ligands.
  JOURNAL   Dev Cell. 2020 Sep 14;54(5):583-592.e5. doi: 
            10.1016/j.devcel.2020.08.004. Epub 2020 Sep 3.
  PUBMED    32888416
REFERENCE   2  (bases 1 to 4242)
  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.devcel.2020.08.004"; journalName: "Dev Cell"; date: 
            "2020-09-14- 14"; volume: "54"; issue: "5"; pages: "583-592.e5"
COMMENT     Sequence Label: pTVGFP
FEATURES             Location/Qualifiers
     source          1..4242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..26
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     18..35
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     19..35
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     42..61
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        42..60
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    184..217
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(406..408,409..411,412..1125)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(406..408,409..411,412..1122)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(451..472)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(712..731)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1059..1080
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1245..1366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1282..1301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1336..1355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1375..1408
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1473..1491)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1512..1528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1512..1528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1525..1547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1536..1552
                     /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(1560..1566,1567..1584,1585..1590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1605..1626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1743..1760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1914..2502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1994..2013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2673..3464,3465..3533))
                     /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     3296..3315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3534..3638)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3664..4119)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3801..3822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4013..4032
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
       61 gcgaattgga gctgaattct tgcatgcaat gcggccgcta ccgcgggcta gcacatatgc
      121 aggtaccgta gtgccccaac tggggtaacc tttgagttct ctcagttggg ggcgtagtgt
      181 accataactt cgtataatgt atgctatacg aagttatcgt acggcccgga tctaattcaa
      241 ttagagacta attcaattag agctaattca attagcgccg gagtataaat agaggcgctt
      301 cgtctacgga gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc
      361 taagcaaata aacaagcgca gctgaacaag ctaaacaatc ggcccatggt gagcaagggc
      421 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      481 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      541 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      601 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      661 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      721 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      781 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      841 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      901 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      961 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1021 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1081 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcgcc gcgactctga
     1141 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1201 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1261 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1321 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttagccc cggtataact
     1381 tcgtataatg tatgctatac gaagttatca ctagtgcgac gtcgattaat taagcacgcg
     1441 tgcaccggta gatctcgtac cagcttttgt tccctttagt gagggttaat ttcgagcttg
     1501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3661 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3721 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3781 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3841 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3901 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3961 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4021 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4081 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     4141 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     4201 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gt
//
