LOCUS       Exported                6855 bp ds-DNA     circular SYN 15-AUG-2025
DEFINITION  10.JW_4-A5.
ACCESSION   10.JW_4-A5
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  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..6855
                     /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..676
                     /label=chicken beta-actin promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.6"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     intron          677..1697
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.5"
                     /note="/match_length=100.4"
                     /note="/other=intron"
     CDS             1695..1751
                     /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      1767..1776
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1773..2489
                     /codon_start=1
                     /label=mVenus
                     /note="pLannotate"
                     /note="/database=fpbase"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1977..1979
                     /label=L
     CDS             2511..3134
                     /codon_start=1
                     /product="https://www.ncbi.nlm.nih.gov/nucleotide/NM_001398
                     595.2"
                     /label=rRim1[20-227]_NM_001398595.2
                     /label=RIMS1_MOUSE (fragment)
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=99.0"
                     /note="/match_length=14.2"
                     /note="/other=CDS"
                     /translation="QELPDLSHLTEEERNIIMAVMDRQKEEEEKEEAMLKCVVRDMAKP
                     AACKTPRNAESQPHQPPLNIFRCVCVPRKPSSEEGGPERDWRLHQQFESYKEQVRKIGE
                     EARRYQGEHKDDAPTCGICHKTKFADGCGHLCSYCRTKFCARCGGRVSLRSNNEDKVVM
                     WVCNLCRKQQEILTKSGAWFFGSGPQQPSQDGTLSDTATGAGSEV"
     CDS             3144..3857
                     /codon_start=1
                     /label=mVenus(L68V)
                     /label=SYFP2
                     /note="pLannotate"
                     /note="/database=fpbase"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=99.6"
                     /note="/other=CDS"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3345..3347
                     /label=V
     polyA_signal    3952..4007
                     /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(4368..4384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4392..4408
                     /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        4416..4446
                     /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    4461..4482
                     /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(4495..4534)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     promoter        4541..4736
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4541..4563
                     /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      4587..4722
                     /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    4742..4876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4742..4863
                     /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(4878..4932)
                     /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   4944..4955
                     /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      5115..5703
                     /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(5874..6734)
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
     promoter        6735..6839
                     /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 cgcgcgccag gcggggcggg
      541 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      601 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      661 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      841 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      901 ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc
      961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga
     1021 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg
     1081 ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct
     1141 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg
     1201 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg
     1261 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg
     1321 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     1381 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     1441 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     1501 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     1561 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     1621 cggcttctgg cgtgtgaccg gcggctctag ctagagcctc tgctaaccat gttcatgcct
     1681 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     1741 gcaaagaatt cgctagcgct accggtgcca ccatggtgag caagggcgag gagctgttca
     1801 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1861 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1921 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     1981 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2041 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2101 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2161 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2221 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     2281 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2341 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     2401 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2461 tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgaggc caagaactgc
     2521 ccgacctgag ccacctgacc gaggaggaga ggaacattat catggcagtg atggaccggc
     2581 agaaggaaga ggaggaaaaa gaagaggcca tgctcaagtg tgttgtcagg gacatggcga
     2641 agcctgctgc ctgcaaaaca ccaagaaatg ctgaaagcca gccccatcaa ccaccactga
     2701 acattttcag atgtgtctgt gttcccagaa agccaagcag cgaagaggga ggcccagaaa
     2761 gagactggag attgcatcaa cagtttgaaa gctacaagga gcaagtgaga aaaatcggag
     2821 aggaagcgag gcgttaccag ggcgagcaca aggatgatgc cccgacgtgt ggaatctgtc
     2881 ataagacaaa gtttgctgat ggatgtggcc atctctgctc ctattgtcgc accaagttct
     2941 gtgcacgctg cggaggccgt gtgtctctgc gatcgaacaa tgaggacaaa gtggttatgt
     3001 gggtatgcaa tttatgtcga aagcaacaag aaatcttaac gaaatctgga gcgtggttct
     3061 ttggaagtgg ccctcagcag cctagtcaag atgggactct gagtgacacg gccacaggtg
     3121 ctggatctga ggtgggatcc ggagtgagca agggcgagga gctgttcacc ggggtggtgc
     3181 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3241 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     3301 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
     3361 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3421 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3481 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3541 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3601 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     3661 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3721 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     3781 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3841 tggacgagct gtataagtaa gcggccgcga ctctagagtc gacggatctt tttccctctg
     3901 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3961 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4021 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4081 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     4141 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     4201 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     4261 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     4321 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     4381 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4441 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4501 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatac gcatctcaat tagtcagcaa
     4561 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4621 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4681 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4741 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4861 ttatcatgtc tcgatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4921 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5521 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6181 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6481 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6841 acatttcccc gaaaa
//
