LOCUS       sequence-258290-        3953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon based transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3953)
  AUTHORS   Stanney W 3rd, Ladam F, Donaldson IJ, Parsons TJ, Maehr R, Bobola N,
            Sagerstrom CG
  TITLE     Combinatorial action of NF-Y and TALE at embryonic enhancers defines
            distinct gene expression programs during zygotic genome activation 
            in zebrafish.
  JOURNAL   Dev Biol. 2020 Mar 15;459(2):161-180. doi: 
            10.1016/j.ydbio.2019.12.003. Epub 2019 Dec 17.
  PUBMED    31862379
REFERENCE   2  (bases 1 to 3953)
  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.ydbio.2019.12.003"; journalName: "Dev Biol"; date: 
            "2020-03-15- 15"; volume: "459"; issue: "2"; pages: "161-180"
COMMENT     Sequence Label: pTransgenesis p3 sv40 minimal promoter:: Katus...
FEATURES             Location/Qualifiers
     source          1..3953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..223
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      74..209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             310..1014
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     polyA_signal    1091..1225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1115..1134)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1169..1188
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1329..1424)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1462..1480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1485..1501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1485..1501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1603..1622)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1614..2423
                     /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(1681..1700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2570..3158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3059..3078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3312..3329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3488..3515)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3607..3693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3742..3764
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3756..3773
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3757..3773
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(3844..3953,1..14))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cttgtacaaa gtggggatct gcgatctgca tctcaattag tcagcaacca tagtcccgcc
       61 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg
      121 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
      181 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg gcattccggt
      241 actgttggta aaggatccac catggcatca atgcagaagc tgatctcaga ggaggacctg
      301 agatccacca tggtgggtga ggatagcgtg ctgatcaccg agaacatgca catgaaactg
      361 tacatggagg gcaccgtgaa cgaccaccac ttcaagtgca catccgaggg cgaaggcaag
      421 ccctacgagg gcacccagac catgaagatc aaggtggtcg agggcggccc tctccccttc
      481 gccttcgaca tcctggctac cagcttcatg tacggcagca aaacctttat caaccacacc
      541 cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagaggatc
      601 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccagaacggc
      661 tgcctcatct acaacgtcaa gatcaacggg gtgaacttcc catccaacgg ccctgtgatg
      721 cagaagaaaa cactcggctg ggaggccagc accgagatgc tgtaccccgc tgacagcggc
      781 ctgagaggcc atagccagat ggccctgaag ctcgtgggcg ggggctacct gcactgctcc
      841 ctcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcttctac
      901 ttcgtggaca ggagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
      961 cacgagatgg ctgtggccag gtactgcgac ctgcctagca aactggggca cagctccgga
     1021 ctcagatctc gagctcaagc ttcgaattct gcagtcgagc ctctagaact atagtgagtc
     1081 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1141 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1201 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1261 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc aattgggccc
     1321 ggtaccccca actttattat acaaagttgg cattataaaa aagcattgct tatcaatttg
     1381 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct ccatggtagc
     1441 gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1501 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1561 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     1621 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     1681 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     1741 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1801 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1861 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     1921 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1981 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2041 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2101 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     2161 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2221 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2281 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2341 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2401 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     2461 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     2521 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3241 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3301 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3361 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3421 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     3481 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     3541 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     3601 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     3661 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
     3721 taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc
     3781 tcgggccctg cagctctaga gctcgaattc tacaggtcac taataccatc taagtagttg
     3841 gttcatagtg actgcatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
     3901 atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac ttt
//
