LOCUS       VB200624-1263ezw        6647 bp    DNA     circular SYN 21-APR-2025
DEFINITION  
ACCESSION   VB200624-1263ezw
VERSION     VB200624-1263ezw
KEYWORDS    .
SOURCE      
  ORGANISM  .
            
FEATURES             Location/Qualifiers
     source          1..6647
     ITR             1..141
                     /application_notes="Allows rescue of virus from recombinant
                     plasmid and replication of the viral genome; this ITR is
                     identical to that of the wild-type AAV2 genome."
                     /description="AAV 5' inverted terminal repeat (functional
                     equivalent of wild-type 5' ITR)"
                     /label="5' ITR"
                     /note="color: #0375bb; direction: RIGHT"
                     /official_designation="5' ITR"
                     /uuid="5f14ec1a645a407dab2725d71bd1c86a"
     Promoter        169..1901
                     /application_notes="Strong promoter."
                     /description="CMV early enhancer fused to modified chicken
                     beta-actin promoter"
                     /full_name="Cytomegalovirus early enhancer fused with
                     chicken beta-actin promoter"
                     /label="CAG"
                     /note="color: #a949ca; direction: RIGHT"
                     /official_designation="CAG"
                     /uuid="420ea94873db443a902370384d17a441"
     Miscellaneous   1926..1959
                     /application_notes="Recognition site of Cre recombinase;
                     can't work with other types of Lox site."
                     /description="Mutated Lox site with two base substitutions
                     of LoxP"
                     /label="Lox2272"
                     /note="color: #946a00; direction: RIGHT"
                     /official_designation="Lox2272"
                     /uuid="a19dd1e203df40eb846e5102df13ffb4"
     Miscellaneous   1992..2025
                     /application_notes="Recognition site of Cre recombinase."
                     /description="Locus of X(cross)-over in P1"
                     /label="LoxP"
                     /note="color: #512bbd; direction: RIGHT"
                     /official_designation="LoxP"
                     /uuid="b48e25a55648467f8c98b3f68faf613f"
     CDS             complement(2038..2757)
                     /application_notes="Commonly used green fluorescent
                     protein; ranked high in brightness, photostability and pH
                     stability among all fluorescent proteins."
                     /description="Enhanced green fluorescent protein; codon
                     optimized based on a variant of wild type GFP from the
                     jellyfish Aequorea victoria"
                     /full_name="EGFP reporter gene"
                     /label="EGFP"
                     /note="color: #6ddaae; direction: LEFT"
                     /official_designation="EGFP"
                     /uuid="14bbdeb46af6422aafbe7a45e8854f81"
     Linker          complement(2758..2823)
                     /application_notes="Causes co-translational cleavage of the
                     encoded polypeptide. Multiple proteins can be made from a
                     polycistronic transcript containing multiple ORFs separated
                     by 2A. P2A and T2A have higher cleavage efficiency compared
                     to other 2As."
                     /description="Self-cleaving 2A peptide from Porcine
                     teschovirus-1"
                     /full_name="Porcine teschovirus-1 2A peptide"
                     /label="P2A"
                     /note="color: #d84e4e; direction: LEFT"
                     /official_designation="P2A"
                     /uuid="a4cf236da25b45e4a9d03c06a25d973a"
     CDS             complement(2824..3528)
                     /application_notes=""
                     /label="mPomc[NM_008895.4](ns)"
                     /note="color: #12f548; direction: LEFT"
                     /official_designation="mPomc[NM_008895.4]"
                     /uuid="ddccdd1a7553400799c203a3c8aa1580"
     Miscellaneous   complement(3529..3534)
                     /application_notes="Facilitates translation initiation of
                     ATG start codon downstream of the Kozak sequence."
                     /description="Kozak translation initiation sequence"
                     /full_name="Kozak"
                     /label="Kozak"
                     /note="color: #1f36a9; direction: LEFT"
                     /official_designation="Kozak"
                     /uuid="ba4f837cfff042e38e7561244b24cfa6"
     Miscellaneous   complement(3541..3574)
                     /application_notes="Recognition site of Cre recombinase;
                     can't work with other types of Lox site."
                     /description="Mutated Lox site with two base substitutions
                     of LoxP"
                     /label="Lox2272"
                     /note="color: #db6901; direction: LEFT"
                     /official_designation="Lox2272"
                     /uuid="991e0551b2034abd9c3ad4e364260ba6"
     Miscellaneous   complement(3607..3640)
                     /application_notes="Recognition site of Cre recombinase."
                     /description="Locus of X(cross)-over in P1"
                     /label="LoxP"
                     /note="color: #800c0c; direction: LEFT"
                     /official_designation="LoxP"
                     /uuid="f4dc79ba31354e87bba6736ab88dbb23"
     Regulatory_Elem 3664^3665
                     /application_notes=""
                     /label="No regulatory element"
                     /note="color: #0c5d90; direction: RIGHT"
                     /official_designation="NM_001111647.2(ns)"
                     /uuid="72e1edd2c1a540b28efbb477521f8696"
     PolyA_signal    3695..3902
                     /application_notes="Allows transcription termination and
                     polyadenylation of mRNA transcribed by Pol II RNA
                     polymerase."
                     /description="Bovine growth hormone polyadenylation signal"
                     /full_name="Bovine growth hormone polyadenylation"
                     /label="BGH pA"
                     /note="color: #f58600; direction: RIGHT"
                     /official_designation="BGH pA"
                     /uuid="4a09734ca6c745b3aadc757c1638b651"
     ITR             complement(3910..4050)
                     /application_notes="Allows rescue of virus from recombinant
                     plasmid and replication of the viral genome; this ITR is
                     identical to that of the wild-type AAV2 genome."
                     /description="AAV 3' inverted terminal repeat"
                     /label="3' ITR"
                     /note="color: #5566f5; direction: LEFT"
                     /official_designation="3' ITR"
                     /uuid="5b8101593b634e64aa9eef9434216f71"
     CDS             4967..5827
                     /application_notes="Allows E. coli to be resistant to
                     ampiciIIin."
                     /description="AmpiciIIin resistance gene"
                     /full_name="Ampicillin resistance gene"
                     /label="AmpiciIIin"
                     /note="color: #ef6cdf; direction: RIGHT"
                     /official_designation="Ampicillin"
                     /uuid="5bcc09480c0e4c07b95ddccd8242d769"
     Rep_origin      5998..6586
                     /application_notes="Facilitates plasmid replication in E.
                     coli; regulates high-copy plasmid number (500-700)."
                     /description="pUC origin of replication"
                     /full_name="pUC origin of replication"
                     /label="pUC ori"
                     /note="color: #c54b7c; direction: RIGHT"
                     /official_designation="pUC ori"
                     /uuid="b978ad4ff785460db448caf63b728237"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgct cgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
      601 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
      661 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
      721 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      781 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      841 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      901 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      961 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
     1021 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
     1081 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
     1141 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1201 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1261 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1321 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1381 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1441 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1501 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1561 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1621 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1681 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1741 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1801 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1861 tgctggttat tgtgctgtct catcattttg gcaaagaatt gcaagtttgt acaaaaaagc
     1921 aggctataac ttcgtatagg atactttata cgaagttatc ctcgagtacc ggtcacgcgt
     1981 acgcaagctt gataacttcg tatagcatac attatacgaa gttatcctag ggccacctta
     2041 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     2101 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     2161 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     2221 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     2281 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     2341 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     2401 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     2461 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     2521 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     2581 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     2641 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     2701 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatagg
     2761 cccggggttt tcttcaacat ctcctgcttg ctttaacaga gagaagttcg tggctccgct
     2821 tccctggccc ttcttgtgcg cgttcttgat gatggcgttc ttgaagagcg tcaccagggg
     2881 cgtctggctc ttctcggagg tcatgaagcc accgtaacgc ttgtccttgg gcgggttgct
     2941 ccagcggaag tgctccaccc ggtagggccc gtcgtccttc tccgcgtcgg actccaggac
     3001 ctgctccaag cctaatggcc gctcgccttc cagctccctc ttgaactcta ggggaaaggc
     3061 ctccgccgac tcgttctcag caacgttggg gtacaccttc accgggcgcc gtttcttgcc
     3121 caccggcttg ccccagcgga agtgctccat ggagtaggag cgcttgccct cgcgtggact
     3181 cggctctgga ctgccatctc cccacaccgc ctcttcctcc gcacgcctct gcgccgcgct
     3241 gccagcactg ctgctgttcc tggggccgaa gcggtcccag cggaagtgac ccatgacgta
     3301 cttccggggg ttttcagtca ggggctgttc atctccgttg ccaggaaaca cgggcgtctc
     3361 cagcgagagg tcgagtttgc aagcccggat gcaagccagc aggttgctct ccgtggtgag
     3421 gtcctggcac tggctgctct ccaggcacca gctccacaca tctatggagg tctgaagcag
     3481 gagggccagc aacagggccc ctgagcgact gtagcagaat ctcggcatgg tggctctaga
     3541 ataacttcgt ataaagtatc ctatacgaag ttataggtac ctgagctcaa catcgatagc
     3601 actagtataa cttcgtataa tgtatgctat acgaagttat acccagcttt cttgtacaaa
     3661 gtgggaattc ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3721 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3781 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3841 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     3901 gagggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3961 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4021 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4081 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4141 cgcattaagc gcggcggggg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4501 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4561 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4621 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4681 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4741 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4801 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4981 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5041 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5101 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5161 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5221 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5281 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5341 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5401 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5461 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5521 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5581 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5641 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     5701 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5761 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5821 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5881 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
