LOCUS       Exported                6165 bp ds-DNA     circular SYN 15-AUG-2025
DEFINITION  12.JW_4-A7.
ACCESSION   12.JW_4-A7
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  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..6165
                     /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..678
                     /label=chicken beta-actin promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.6"
                     /note="/match_length=100.4"
                     /note="/other=promoter"
     intron          679..1699
                     /label=chimeric intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.5"
                     /note="/match_length=100.4"
                     /note="/other=intron"
     CDS             1697..1753
                     /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      1770..1779
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1776..2492
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2514..3167
                     /codon_start=1
                     /product="https://www.ncbi.nlm.nih.gov/nucleotide/NM_013019
                     .2"
                     /label=rRab4a(S27N)_NM_013019.2
                     /label=RAB4A_RAT
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=99.5"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MAQTAMSETYDFLFKFLVIGNAGTGKNCLLHQFIEKKFKDDSNHT
                     IGVEFGSKIINVGGKYVKLQIWDTAGQERFRSVTRSYYRGAAGALLVYDITSRETYNAL
                     TNWLTDARMLASQNIVIILCGNKKDLDADREVTFLEASRFAQENELMFLETSALTGENV
                     EEAFMQCARKILNKIESGELDPERMGSGIQYGDAALRQLRSPRRTQAPSAQECGC"
     misc_feature    2592..2594
                     /label=27N
     polyA_signal    3262..3317
                     /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(3678..3694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3702..3718
                     /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        3726..3756
                     /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    3771..3792
                     /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(3805..3844)
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=3.7"
                     /note="/other=CDS"
     promoter        3851..4046
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3851..3873
                     /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      3897..4032
                     /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    4052..4186
                     /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(4188..4242)
                     /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   4254..4265
                     /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      4425..5013
                     /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(5184..6044)
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
     promoter        6045..6149
                     /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 ccccccctcc ccacccccaa ttttgtattt atttattttt
      481 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      541 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      601 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      661 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      721 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      781 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      841 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      901 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
      961 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     1021 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     1081 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     1141 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     1201 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     1261 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     1321 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     1381 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     1441 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     1501 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     1561 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1621 ttcggcttct ggcgtgtgac cggcggctct agctagagcc tctgctaacc atgttcatgc
     1681 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1741 tggcaaagaa ttcgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     1801 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1861 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1921 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     1981 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2041 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2101 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2161 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2221 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2281 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2341 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     2401 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2461 ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga gggatggcgc
     2521 agaccgccat gtccgagact tacgatttct tgtttaagtt cttggtcatt ggaaatgcgg
     2581 gaactggcaa aaactgcttg ctccatcagt tcattgagaa gaaattcaaa gatgactcaa
     2641 atcataccat aggagtggaa ttcggctcaa agataataaa tgttggtggt aaatatgtga
     2701 agttacagat atgggacacg gctggacagg agcggttcag gtctgtgacg agaagctact
     2761 acagaggtgc ggctggggca ctcctcgtct atgacatcac cagccgagaa acctacaatg
     2821 cgcttactaa ttggttaaca gatgccagaa tgctggcgag ccagaacatc gtcatcattc
     2881 tctgcgggaa caagaaggac ctggatgccg accgggaagt caccttcctt gaagcctcca
     2941 ggttcgcaca agagaatgag ctcatgttcc tggaaaccag tgcactgact ggcgagaacg
     3001 tcgaagaggc tttcatgcag tgcgcaagga agatacttaa caaaattgaa tcaggtgagc
     3061 tggaccccga gaggatgggc tctggtatcc agtatggaga cgccgccttg agacagctac
     3121 ggtcaccccg acgtacacag gctccaagtg cacaggagtg tggctgctag gcggccgcga
     3181 ctctagagtc gacggatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3241 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3301 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     3361 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     3421 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     3481 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3541 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3601 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     3661 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3721 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3781 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3841 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3961 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4021 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     4081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     4201 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4261 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4321 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4381 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4441 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4501 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4561 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4621 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4681 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4741 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4801 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4861 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4921 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4981 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5041 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5101 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5161 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5221 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5281 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5341 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5401 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5461 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5521 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5581 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5641 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5701 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5761 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5821 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5881 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5941 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6001 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6061 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6121 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaa
//
