LOCUS       pAAV-EF1a-DIO-JEDI-1P-W 6898 bp ds-DNA     circular     16-MAY-2023
DEFINITION  Cre-activated AAV expression of JEDI-1P.
ACCESSION   urn.local...1c-bb4p8kc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct .
COMMENT     Imported using the Genbank importer. File name:
            2298_pAAV-EF1a-DIO-JEDI-1P-WPRE (modified).gb
FEATURES             Location/Qualifiers
     repeat_region   2..131
                     /label="AAV2 ITR (alternate)"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="note: Functional equivalent of wild-type AAV2 ITR"
     misc_feature    3..8
                     /label="PstI"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="Recognition_pattern: CTGCA^G annotation_group: CTGCAG note: Geneious type: restriction site"
     misc_feature    41..46
                     /label="SmaI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="Recognition_pattern: CCC^GGG annotation_group: CCCGGG note: Geneious type: restriction site"
     misc_feature    132..139
                     /label="NotI"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="Recognition_pattern: GC^GGCCGC annotation_group: GCGGCCGC note: Geneious type: restriction site"
     misc_feature    146..151
                     /label="HindIII"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="Recognition_pattern: A^AGCTT annotation_group: AAGCTT note: Geneious type: restriction site"
     promoter        232..1410
                     /label="EF-1-alpha promoter"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="note: strong constitutive promoter for human elongation factor EF-1-alpha"
     intron          463..1401
                     /label="EF-1-alpha intron A"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="note: intron upstream of the start codon of human EF-1-alpha"
     misc_feature    544..549
                     /label="PstI"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="Recognition_pattern: CTGCA^G annotation_group: CTGCAG note: Geneious type: restriction site"
     misc_feature    611..616
                     /label="AflII"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /note="Recognition_pattern: C^TTAAG annotation_group: CTTAAG note: Geneious type: restriction site"
     misc_feature    798..803
                     /label="BglII"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="Recognition_pattern: A^GATCT annotation_group: AGATCT note: Geneious type: restriction site"
     misc_feature    1049..1054
                     /label="PstI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="Recognition_pattern: CTGCA^G annotation_group: CTGCAG note: Geneious type: restriction site"
     misc_feature    1197..1202
                     /label="XhoI"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="Recognition_pattern: C^TCGAG annotation_group: CTCGAG note: Geneious type: restriction site"
     primer_bind     1358..1378
                     /label="EF-1aForward"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 28443813537970036b9e755-8eaa-4066-8402-41d62495d47e %GC: 52.4 Tm: 59.9 Hairpin_Tm: 33.4 Self_Dimer_Tm: None Sequence: TCAAGCCTCAGACAGTGGTTC"
     misc_feature    1411..1416
                     /label="KpnI"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="Recognition_pattern: GGTAC^C annotation_group: GGTACC note: Geneious type: restriction site"
     misc_feature    1417..1422
                     /label="BamHI"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="Recognition_pattern: G^GATCC annotation_group: GGATCC note: Geneious type: restriction site"
     misc_feature    1417..1422
                     /label="BamHI"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="Recognition_pattern: G^GATCC annotation_group: GGATCC note: Geneious type: restriction site"
     misc_feature    1423..1428
                     /label="XbaI"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="Recognition_pattern: T^CTAGA annotation_group: TCTAGA note: Geneious type: restriction site"
     misc_feature    1429..1434
                     /label="SalI"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="Recognition_pattern: G^TCGAC annotation_group: GTCGAC note: Geneious type: restriction site"
     misc_feature    1435..1440
                     /label="Kpn2I"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="Recognition_pattern: T^CCGGA annotation_group: TCCGGA note: Geneious type: restriction site"
     protein_bind    1441..1474
                     /label="lox2272"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="bound_moiety: Cre recombinase note: Cre-mediated recombination occurs in the 8-bp coresequence (GGATACTT). lox2272 sites are compatible witheachother, but incompatible with loxP or loxN sites."
     protein_bind    1525..1558
                     /label="loxP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="bound_moiety: Cre recombinase note: Cre-mediated recombination occurs in the 8-bp coresequence (GCATACAT)."
     misc_feature    1575..1580
                     /label="Bsp1407I"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="Recognition_pattern: T^GTACA annotation_group: TGTACA note: Geneious type: restriction site"
     CDS             complement(1587..2867)
                     /label="JEDI-1P"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="created_by: User Transferred_From: pCS2-Hygro-ChickenVSD(R154Q)-Mermaid(partial) Transferred_Similarity: 100.00% modified_by: User modified_by: User modified_by: User"
     CDS             complement(1587..2867)
                     /label="JEDI-1P"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="created_by: User Transferred_From: pCS2-Hygro-ChickenVSD(R154Q)-Mermaid(partial) Transferred_Similarity: 100.00% modified_by: User modified_by: User modified_by: User"
                     /translation="METTVRYEQGSELTKTSSSPTADEPTIKIDDGRDEGNEQDSCSNTIRRKISPFVMSFGFRVFGVVLIIVDIIVVIVDLAISEKKRGIREILEGVSLAIALFFLVDVLMRVFVEGFKNYFRSKLVTLDAVIVVGTLLINMTYSFSDLAAFNSENVYITADKQKNGIKANFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYGGTGGSASQGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGKYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNTDQMPHMVTLLRVLKIVILIRIFRLASQKKQLEVVT"
     modified_base   complement(1650..1652)
                     /label="R406K AAA"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="created_by: User"
     modified_base   complement(1677..1679)
                     /label="Q397H CAC"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="created_by: User"
     misc_feature    complement(1893..1898)
                     /label="NdeI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="Recognition_pattern: CA^TATG annotation_group: CATATG note: Geneious type: restriction site"
     misc_feature    complement(2263..2282)
                     /label="519_cpsfGFP-SB_NT_F"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="Hairpin: 5.0 Primer-Dimer: 1.0 Monovalent_Salt_Concentration: 50.0 Divalent_Salt_Concentration: 0.0 DNTP_Concentration: 0.0 Mismatches: 0 created_by: primer3 annotation_group: 354575745130088644ca8504-1631-4db6-8586-dee58954597d %GC: 65.0 Tm: 65.3 Hairpin_Tm: 42.1 Self_Dimer_Tm: 34.7 Sequence: CCGGTGCTGCTGCCGGATAA note: Geneious type: primer_bind_reverse"
     modified_base   complement(2412..2414)
                     /label="H152E GAG"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="created_by: User"
     modified_base   complement(2496..2498)
                     /label="N124V GTT"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="created_by: User"
     misc_feature    2878..2883
                     /label="NheI"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="Recognition_pattern: G^CTAGC annotation_group: GCTAGC note: Geneious type: restriction site"
     misc_feature    2878..2883
                     /label="NheI"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /note="Recognition_pattern: G^CTAGC annotation_group: GCTAGC note: Geneious type: restriction site"
     protein_bind    complement(2884..2917)
                     /label="lox2272"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="bound_moiety: Cre recombinase note: Cre-mediated recombination occurs in the 8-bp coresequence (GGATACTT). lox2272 sites are compatible witheachother, but incompatible with loxP or loxN sites."
     protein_bind    complement(2968..3001)
                     /label="loxP"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="bound_moiety: Cre recombinase note: Cre-mediated recombination occurs in the 8-bp coresequence (GCATACAT)."
     misc_feature    3002..3007
                     /label="EcoRI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="Recognition_pattern: G^AATTC annotation_group: GAATTC note: Geneious type: restriction site"
     misc_feature    3008..3013
                     /label="EcoRV, Eco32I"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="Recognition_pattern: GAT^ATC annotation_group: GATATC note: Geneious type: restriction site"
     misc_feature    3014..3019
                     /label="HindIII"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="Recognition_pattern: A^AGCTT annotation_group: AAGCTT note: Geneious type: restriction site"
     misc_feature    3026..3614
                     /label="WPRE"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="note: woodchuck hepatitis virus posttranscriptional regulatory element"
     CDS             complement(3497..3508)
                     /label="Factor Xa site"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="product: Factor Xa recognition and cleavage site translation: IEGR"
     CDS             complement(3497..3508)
                     /label="Factor Xa site"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="product: Factor Xa recognition and cleavage site translation: IEGR"
                     /translation="IEGR"
     misc_feature    3632..3637
                     /label="XhoI"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="Recognition_pattern: C^TCGAG annotation_group: CTCGAG note: Geneious type: restriction site"
     misc_feature    3638..3643
                     /label="BglII"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="Recognition_pattern: A^GATCT annotation_group: AGATCT note: Geneious type: restriction site"
     polyA_signal    3646..4122
                     /label="hGH poly(A) signal"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="note: human growth hormone polyadenylation signal"
     misc_feature    complement(3709..3728)
                     /label="XL39"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 2844378089681004f549c5b-6b8d-40fb-8ad5-8557c63b43e5 %GC: 55.0 Tm: 59.4 Hairpin_Tm: None Self_Dimer_Tm: None Sequence: ATTAGGACAAGGCTGGTGGG note: Geneious type: primer_bind_reverse"
     misc_feature    4154..4161
                     /label="NotI"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /note="Recognition_pattern: GC^GGCCGC annotation_group: GCGGCCGC note: Geneious type: restriction site"
     repeat_region   4162..4291
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     repeat_region   4162..4302
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: inverted terminal repeat of adeno-associated virusserotype 2"
     misc_feature    4247..4252
                     /label="SmaI"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="Recognition_pattern: CCC^GGG annotation_group: CCCGGG note: Geneious type: restriction site"
     misc_feature    4258..4263
                     /label="SmaI"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="Recognition_pattern: CCC^GGG annotation_group: CCCGGG note: Geneious type: restriction site"
     misc_feature    4296..4301
                     /label="PstI"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="Recognition_pattern: CTGCA^G annotation_group: CTGCAG note: Geneious type: restriction site"
     rep_origin      4377..4832
                     /label="f1 ori"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="direction: RIGHT note: f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
     primer_bind     4674..4695
                     /label="F1ori-F"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="note: F1 origin, forward primer"
     primer_bind     complement(4849..4868)
                     /label="pRS-marker"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: pRS vectors, use to sequence yeast selectable marker"
     misc_feature    complement(4914..4934)
                     /label="pexF"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 284438067391300ce63af36-3c3a-4047-b84e-bce1b23d2421 %GC: 66.7 Tm: 65.3 Hairpin_Tm: 38.4 Self_Dimer_Tm: None Sequence: GGAGCAGACAAGCCCGTCAGG note: Geneious type: primer_bind_reverse"
     misc_feature    complement(4914..4934)
                     /label="pexF"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 215713782318200433134159-0716-4bc7-b149-3672b918d8b3 %GC: 66.7 Tm: 65.3 Hairpin_Tm: 38.4 Self_Dimer_Tm: None Sequence: GGAGCAGACAAGCCCGTCAGG note: Geneious type: primer_bind_reverse"
     primer_bind     4968..4990
                     /label="3GEX"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 284405544235300d122ac66-812b-41c5-b19f-af44fbeede05 %GC: 65.2 Tm: 67.3 Hairpin_Tm: None Self_Dimer_Tm: 1.6 Sequence: CCGGGAGCTGCATGTGTCAGAGG"
     primer_bind     4972..4990
                     /label="pGexR"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 284438003260600823a9de3-785a-4d7b-83bb-1a025aa07584 %GC: 57.9 Tm: 58.2 Hairpin_Tm: None Self_Dimer_Tm: 8.1 Sequence: GAGCTGCATGTGTCAGAGG"
     misc_feature    complement(5018..5039)
                     /label="pQEfPR"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 28443793817820023c397b6-7442-44a5-919c-4fbe5147cb43 %GC: 54.5 Tm: 62.1 Hairpin_Tm: 47.4 Self_Dimer_Tm: 8.7 Sequence: GTATCACGAGGCCCTTTCGTCT note: Geneious type: primer_bind_reverse"
     misc_feature    complement(5018..5039)
                     /label="pQEfPR"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 35455238429034006d35a26e-d58e-44f5-b061-57cc16204e87 %GC: 54.5 Tm: 62.1 Hairpin_Tm: 47.4 Self_Dimer_Tm: 8.7 Sequence: GTATCACGAGGCCCTTTCGTCT note: Geneious type: primer_bind_reverse"
     primer_bind     complement(5028..5046)
                     /label="pBRforEco"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="note: pBR322 vectors, upsteam of EcoRI site, forward primer"
     misc_feature    complement(5077..5096)
                     /label="VF2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 284437873894500f500c32a-9c1e-4914-a4d3-b19d2987fc1c %GC: 50.0 Tm: 58.7 Hairpin_Tm: None Self_Dimer_Tm: 2.3 Sequence: TGCCACCTGACGTCTAAGAA note: Geneious type: primer_bind_reverse"
     misc_feature    complement(5077..5096)
                     /label="VF2"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="Mismatches: 0 created_by: primer3 annotation_group: 2157137421479433e05f7087-8cc2-4b1c-a3e3-99baeafe48f0 %GC: 50.0 Tm: 58.7 Hairpin_Tm: None Self_Dimer_Tm: 2.3 Sequence: TGCCACCTGACGTCTAAGAA note: Geneious type: primer_bind_reverse"
     promoter        5114..5218
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="gene: bla"
     CDS             5219..6079
                     /label="AmpR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="gene: bla product: beta-lactamase note: confers resistance to ampicillin, carbenicillin,and related antibiotics translation: MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             5219..6079
                     /label="AmpR"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="gene: bla product: beta-lactamase note: confers resistance to ampicillin, carbenicillin,and related antibiotics translation: MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     complement(5437..5456)
                     /label="Amp-R"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="note: Ampicillin resistance gene, reverse primer"
     rep_origin      6250..6838
                     /label="ori"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="direction: RIGHT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
      181 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      241 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      301 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      361 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      421 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      481 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      541 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      601 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      661 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      721 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      781 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      841 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      901 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      961 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1021 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1081 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1141 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1201 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1261 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1321 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1381 agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt cgactccgga
     1441 ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg attgtagctg
     1501 ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac gaagttatgg
     1561 cgcgccctat tacttgtaca tcattaggtt accacttcaa gttgtttctt ctgtgaagcc
     1621 aggcgaaata ttcttattaa gataacaatt ttcagaactc gaagaagagt aaccatgtgc
     1681 ggcatctgat ctgtgttata ttccagttta tgccccagaa tgttgccatc ttctttaaaa
     1741 tcggtgcctt tcagttcaat gcggttcacc agggtatcgc cttcaaattt caccaccgcg
     1801 cgggttttat atttgccatc atctttaaag ctaatggtgc gttcctgcac atagccttcc
     1861 ggcatcgcgc ttttaaaaaa atcatggcgt ttcatatgat ccggatagcg gctaaagcac
     1921 tgcacgccat aggttaaggt ggtcaccagg gtcggccacg gcaccggtag tttgccggtg
     1981 gtgcaaataa atttcagggt cagtttgcca atggtcgcat cgccttcgcc ttcgccgcgc
     2041 acgctaaatt tatggccgtt cacatcgcca tccagttcca ccagaatcgg caccacgccg
     2101 gtaaacagtt cttcgccctg gctcgcgctg ccgccggtgc cgccatacag ttcatccatg
     2161 ccgtgtgtaa tgcccgctgc ggtcacaaat tccagcagca ccatgtgatc gcgtttttcg
     2221 ttcggatctt tgctcagcac ggtctgggtg ctcagataat ggttatccgg cagcagcacc
     2281 gggccatcgc caatcggggt gttctgctga taatgatccg ccagctgcac gctgccatct
     2341 tccacgttat ggcgcacggt aaagttcgct ttaatgccgt ttttctgttt atccgcggta
     2401 atatacacgt tctcgctgtt aaaggcagca aggtcagaga aggagtaggt catattaatt
     2461 agcagagtgc ccactactat gactgcatcc aaagtaacca gtttggaccg gaaatagttc
     2521 ttgaagcctt caacaaacac tctcatgaga acatcaacaa ggaagaagag tgctatagcc
     2581 agggaaacac cttcaagaat ctctctaatg cctcttttct tctcactgat ggccagatcc
     2641 acaatcacca ctatgatgtc tacaatgata agcacaactc caaatactct gaatccaaat
     2701 gacatcacaa acggggaaat ttttctccta atggtattgg aacagctgtc ttgttcatta
     2761 ccctcatcac gaccatcatc aatctttatc gtgggctcat ctgctgttgg agagctcgaa
     2821 gttttagtga gctctgaccc ctgttcatac ctcacagtcg tctccatggt ggcggtagct
     2881 agcataactt cgtataaagt atcctatacg aagttatttg ccttaaccca gaaattatca
     2941 ctgttattct ttagaatggt gcaaagaata acttcgtata atgtatgcta tacgaagtta
     3001 tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3061 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3121 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3181 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3241 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3301 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3361 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3421 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     3481 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3541 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3601 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     3661 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3721 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     3781 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     3841 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     3901 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     3961 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4021 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4081 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     4141 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4201 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4261 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4321 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4381 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4441 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4501 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4561 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4621 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4681 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4741 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4801 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     4861 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4921 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4981 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5041 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5101 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5161 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5221 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5281 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5341 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5401 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5461 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5521 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5581 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5641 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5701 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5761 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5821 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5881 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5941 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6001 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6061 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6121 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6181 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6241 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6301 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6361 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6421 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6481 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6541 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6601 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6661 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6721 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6781 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6841 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatg
//