LOCUS       Exported                3589 bp ds-DNA     circular SYN 16-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3589)
  AUTHORS   Baylor-NRI
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3589
                     /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"
     misc_feature    complement(481..498)
                     /label=SeqLa
     misc_feature    537..553
                     /label=M13 For -20
     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=oJDW 2128
     regulatory      700..709
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             join(706..1198,1457..1685)
                     /codon_start=1
                     /label=EGFP
                     /label=EGFP 5' half
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VKLQDPPQHRGRQRAARRPLPAEHPHRRRPRAAARQPLPEHPVRPEQRPQREARSHGPA
                     GVRDRRRDHSRHGRAVQV"
     intron          1199..1456
                     /label=Artificial intron and MCS
     intron          1199..1274
                     /label=Intron Total
     misc_feature    1271..1413
                     /label=Homology to Human HPRT
     primer_bind     complement(1672..1735)
                     /label=oJDW 2129
     protein_bind    complement(1714..1813)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1831..1849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1854..1870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1884..1905
                     /label=SeqL B
     CDS             1983..2792
                     /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      2939..3527
                     /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 caacacattg 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 ccgaattcca tgatcttttt tttttttttt tttttcccat gatcatcccg
     1261 gggatcctgc agatgcagtc agatttctga actgtctcag caaatggtaa gtaacttaga
     1321 gctagtaatt aataacctgt cctttgattt ctgattcagc caagaatggc catatttggg
     1381 aaaggcagat ctggagagta accacgtttt catgcagtct agactgttaa cgcagtgttt
     1441 ctctaactct caggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1561 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1681 gtaaagcggc cgcgactcta gacgcgtact agtgcccagc tttcttgtac aaagttggca
     1741 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     1801 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     1861 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     1921 aaataatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1981 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2041 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2101 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2161 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2221 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2281 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2341 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2401 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2461 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2521 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2581 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2641 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2701 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2761 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     2821 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     2881 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2941 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3001 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3061 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3121 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3181 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3241 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3301 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3361 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3421 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3481 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3541 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
