LOCUS       Exported                4107 bp ds-DNA     circular SYN 09-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4107
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     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"
     primer_bind     645..692
                     /label=Primer 1
     misc_feature    669..691
                     /label=MCS (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=28.4"
                     /note="/other=misc_feature"
     CDS             706..1198
                     /codon_start=1
                     /label=EGFP
                     /label=EGFP 5' half
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     V"
     regulatory      706..709
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     intron          1199..1218
                     /label=Artificial intron and MCS
     misc_feature    1225..1724
                     /label=human Chr 9 / EGFL7
     ncRNA           1460..1532
                     /label=mir-126
                     /note="pLannotate"
                     /note="/database=Rfam"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=ncRNA"
     intron          1725..1974
                     /label=Artificial intron and MCS
     misc_feature    1743..1763
                     /label=critical T stretch
     CDS             1975..2203
                     /codon_start=1
                     /label=EGFP
                     /label=EGFP 5' half
                     /translation="LQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQ
                     RPQREARSHGPAGVRDRRRDHSRHGRAVQV"
     primer_bind     complement(2190..2253)
                     /label=Primer 2
     protein_bind    complement(2232..2331)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2501..3310
                     /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"
     rep_origin      3457..4045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 agcaggcagc tagcgctacc ggactcagat ccaccggtcg ccaccatggt gagcaagggc
      721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      901 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1141 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgagt
     1201 ggatcgggta ccgaattcgc ccttagtgga cattgccgtg tggctgttag gcatggtggg
     1261 gggcactgga atctgggcgg aaggcggtgg ggactccctc tccagggagg gaggatgggg
     1321 agggaggata ggtgggttcc cgagaactgg gggcaggttg cccggagcct catatcagcc
     1381 aagaaggcag aagtgccccg tcccggggtc ctgtctgcat ccagcgcagc attctggaag
     1441 acgccacgcc tccgctggcg acgggacatt attacttttg gtacgcgctg cgacacttca
     1501 aactcgtacc gtgagtaata atgcgccgtc cacggcaccg catcgaaaac gccgctgaga
     1561 cctcagcctt gacctccctc agcgtggccg ggaccctgag cctctgcgca gagccacccg
     1621 ccccgacgta cttaggcggc atagccctga gacctctggc cagcgccagg caggcagcgg
     1681 gggcggcaga ggcctgggcc tgagtcttct ggctctgcct ctccaagggc gaattccatg
     1741 atcttttttt tttttttttt tttcccatga tcatcccggg gatcctgcag atgcagtcag
     1801 atttctgaac tgtctcagca aatggtaagt aacttagagc tagtaattaa taacctgtcc
     1861 tttgatttct gattcagcca agaatggcca tatttgggaa aggcagatct ggagagtaac
     1921 cacgttttca tgcagtctag actgttaacg cagtgtttct ctaactctca ggtgaacttc
     1981 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2041 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2101 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2161 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     2221 cgcgtactag tgcccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2281 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2341 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2401 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2461 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2521 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2581 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2641 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2701 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2761 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2821 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2881 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2941 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3001 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3061 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3121 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3181 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3241 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3301 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3361 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4081 cttttgctgg ccttttgctc acatgtt
//
