LOCUS       sequence-476607-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human DDX6 fused to EGFP in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  TITLE     Su lab plasmids 2025
REFERENCE   2  (bases 1 to 5316)
  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"
COMMENT     Sequence Label: PFN29A-DDX6
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             76..93
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             106..123
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(1591..1593,1594..1596,1597..2310)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1636..1657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1897..1916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2244..2265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2420..2438)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2434..2481
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2710..2814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2815..2883,2884..3675)
                     /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     complement(3033..3052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4537..4820
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     primer_bind     complement(4880..4897)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5050..5136
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5228..5255
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg tattttaact ttactaagga
       61 gaattcatca tgaaacatca tcaccatcac cacgagccaa ccactctggt tccgcgtgga
      121 tccgataacg cgagcacggc cagaacagag aaccctgtta taatgggtct gtccagtcaa
      181 aatggtcagc tgagaggccc tgtgaaaccc actggtggcc ctggaggagg gggcacacag
      241 acacagcaac agatgaacca gctgaaaaac accaacacaa tcaataatgg cactcagcag
      301 caagcacaga gtatgaccac cactattaaa cctggtgatg actggaaaaa gactttaaaa
      361 ctccctccaa aggatctaag aatcaaaact tcggatgtga cctccacaaa aggaaatgag
      421 tttgaagatt actgtttgaa acgggagtta ctgatgggaa tttttgaaat gggctgggaa
      481 aagccatctc ctattcagga ggagagcatt cccattgctt tatctggtag ggatatctta
      541 gctagagcaa aaaatggaac aggcaagagc ggtgcctacc tcattccctt acttgaacgg
      601 ctagacctga agaaggacaa tatacaagca atggtgattg ttcccactag agaacttgct
      661 ctacaggtca gtcaaatttg catccaggtc agcaaacaca tgggaggggc caaagtgatg
      721 gcaaccacag gaggaaccaa tttacgagat gacataatga ggcttgatga tacagtgcac
      781 gtggtgattg ctacccctgg gagaatcctg gatcttatta agaaaggagt agcaaaggtt
      841 gatcatgtcc agatgatagt attggatgag gcagataagt tgctgtcaca ggattttgtg
      901 cagataatgg aggatattat tctcacgcta cctaaaaaca ggcagatttt actatattcc
      961 gctactttcc ctcttagtgt acagaagttc atgaattccc atttgcagaa accctatgag
     1021 attaacctga tggaggaact aactctgaag ggagtaaccc agtactacgc atatgtaact
     1081 gagcgccaaa aagtacactg cctcaacaca cttttctcca ggcttcagat aaaccagtcg
     1141 atcattttct gtaactcctc tcagcgagtt gaattgctag ccaagaagat ttctcaactg
     1201 ggttattctt gcttctatat tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt
     1261 catgatttcc gaaatggctt atgccgcaat cttgtttgca ctgatctgtt tacccgaggt
     1321 attgatatac aagctgtgaa tgtggtaata aactttgatt tcccaaagct ggcagagacc
     1381 tatctccatc gtattggaag atcaggtcgc tttggtcatc ttggcttagc catcaacttg
     1441 atcacatatg atgatcgctt caacctgaaa agtattgagg agcagctggg aacagaaatt
     1501 aaacctattc cgagcaacat tgataagagc ctgtatgtgg cagaatacca cagcgagcct
     1561 gtagaagatg agaaacctga gccaaccact atggtgagca agggcgagga gctgttcacc
     1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1681 tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1801 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2041 gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
     2101 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2161 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2281 actctcggca tggacgagct gtacaagtaa aaacgaattc gggctcggta cccggggatc
     2341 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     2401 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     2461 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggttcgct tgctgtccat
     2521 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     2581 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     2641 cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg
     2701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3001 cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac tatcggagga
     3241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3661 attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg
     4561 gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg
     4621 gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg
     4681 cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat
     4741 cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg
     4801 tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag
     4861 aggatcgaga tcttttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     4921 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     4981 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     5041 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     5221 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     5281 atttttctaa atacattcaa atatgtatcc gctcat
//
