LOCUS       Exported                6950 bp ds-DNA     circular SYN 15-AUG-2025
DEFINITION  06.JW_3-E3.
ACCESSION   06.JW_3-E3
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 21, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.2
FEATURES             Location/Qualifiers
     source          1..6950
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=enhancer"
     promoter        400..677
                     /label=chicken beta-actin promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     intron          678..1698
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.5"
                     /note="/match_length=100.4"
                     /note="/other=intron"
     CDS             1696..1752
                     /label=HBB_LEPEU (fragment)
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=94.7"
                     /note="/match_length=12.9"
                     /note="/other=CDS"
     regulatory      1769..1778
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1775..2482
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             2504..3235
                     /codon_start=1
                     /product="https://www.ncbi.nlm.nih.gov/nucleotide/NM_001433
                     268.1"
                     /label=rFYCO1(963-1206)_NM_001433268.1
                     /label=FYCO1_MOUSE (fragment)
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=89.1"
                     /note="/match_length=17.5"
                     /note="/other=CDS"
                     /translation="QSLQEAAHQEQDALKFQLSAEIMDHQNRLKTANEECGSLRAQLEE
                     QGQELQMTKEAVQELKITKATMEEKLNCTGSHLAECQATLLRKDEESAMLRINLERTQK
                     ELEKATSKFQEYYNKLCQEVTNRERNDQKMLADLDDLNRTKKYLEERLIELLRDKDALW
                     QKSDALEFQQKLSAEEKCLGDMEANHCHDCKREFSWIVRRHHCRLCGRIFCYYCCNNYV
                     VTKPSGKKERCCRACFQKFGES"
     CDS             3245..3955
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    4047..4102
                     /label=beta-globin poly(A) signal
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=polyA_signal"
     primer_bind     complement(4463..4479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4487..4503
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        4511..4541
                     /label=lac promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    4556..4577
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     CDS             complement(4590..4629)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     promoter        4636..4831
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4636..4658
                     /label=SV40 promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=7.7"
                     /note="/other=promoter"
     rep_origin      4682..4817
                     /direction=RIGHT
                     /label=SV40 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     polyA_signal    4837..4971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4837..4958
                     /label=SV40 poly(A) signal
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=polyA_signal"
     CDS             complement(4973..5027)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=5.1"
                     /note="/other=CDS"
     repeat_region   5039..5050
                     /label=AAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=itrs"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.3"
                     /note="/other=repeat_region"
     rep_origin      5210..5798
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             complement(5969..6829)
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
     promoter        6830..6934
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
ORIGIN
        1 gtgccacctg ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      421 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      481 aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg
      541 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      601 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      661 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      721 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      781 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      841 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
      901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
      961 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1021 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1081 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1141 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1201 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1261 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     1321 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1381 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1441 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1501 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1561 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1621 tcggcttctg gcgtgtgacc ggcggctcta gctagagcct ctgctaacca tgttcatgcc
     1681 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     1741 ggcaaagaat tcgctagcgc taccggtcgc caccatggtg agcaagggcg aggaggacaa
     1801 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1861 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1921 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca
     1981 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     2041 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2101 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     2161 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2221 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     2281 gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct acaaggccaa
     2341 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagctggaca tcacctccca
     2401 caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc actccaccgg
     2461 cggcatggac gagctgtaca agtccggact cagatctcga ggccagagcc tccaagaggc
     2521 tgcacaccag gaacaagatg cccttaagtt ccagctaagt gctgagatta tggaccacca
     2581 gaacagacta aagacagcca atgaagagtg tgggagcctc agggcccagc tagaagaaca
     2641 aggccaggag ctgcaaatga ctaaggaggc tgtgcaggaa ctgaagatca ccaaggcgac
     2701 catggaggag aagctgaatt gcaccggtag ccaccttgca gagtgccagg ccactttgct
     2761 acgcaaagat gaagaaagcg ctatgcttcg aatcaaccta gagagaacac agaaggaact
     2821 tgaaaaggct acatcaaaat tccaagaata ttacaacaaa ctctgccagg aggtgacaaa
     2881 cagggaaagg aatgaccaga agatgcttgc agacctagat gacctgaaca gaaccaagaa
     2941 ataccttgag gagcggctga tagagctgct cagggacaaa gatgctctct ggcaaaagtc
     3001 cgatgcactg gaattccagc agaagctcag tgctgaagag aaatgtcttg gggacatgga
     3061 ggccaaccac tgccatgact gcaagcggga gttcagctgg atagtgcggc ggcaccactg
     3121 caggttatgt ggccgaatat tctgttacta ctgctgtaac aactatgttg tgactaagcc
     3181 cagtggcaag aaggaacggt gctgccgagc ctgcttccag aagtttggtg aaagcggatc
     3241 cggaatggtg agcaagggcg aggaggacaa catggccatc atcaaggagt tcatgcgctt
     3301 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     3361 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
     3421 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     3481 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     3541 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     3601 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     3661 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     3721 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     3781 cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     3841 cgtcaacatc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     3901 cgagcgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagcggc
     3961 cgcgactcta gagtcgacgg atctttttcc ctctgccaaa aattatgggg acatcatgaa
     4021 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     4081 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     4141 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     4201 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     4261 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     4321 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     4381 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     4441 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4501 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4561 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4621 cgtgccagcg gatacgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4681 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4741 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4801 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     4861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4921 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctcgat ccgctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
//
