LOCUS       pAttBFGNeo2             6222 bp    DNA        circular     31-JUL-2025
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     vec 3 PCR from 8 (BsaI) to 2394 (BsaI) 
COMMENT     neo 3 PCR from 8 (BsaI) to 3842 (BsaI) 
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     rep_origin      complement(1121..1427)
                     /locus_tag="F1 ori"
                     /label="F1 ori"
                     /ApEinfo_label="F1 ori"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(1..858)
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(1446..1514)
                     /locus_tag="LacZ alpha"
                     /label="LacZ alpha"
                     /ApEinfo_label="LacZ alpha"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(5438..6125)
                     /locus_tag="ColE1 origin"
                     /label="ColE1 origin"
                     /ApEinfo_label="ColE1 origin"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(859..963)
                     /locus_tag="AmpR promoter"
                     /label="AmpR promoter"
                     /ApEinfo_label="AmpR promoter"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_recomb     2197..2260
                     /locus_tag="attB phiC31"
                     /label="attB phiC31"
                     /ApEinfo_label="attB phiC31"
                     /ApEinfo_fwdcolor="#929000"
                     /ApEinfo_revcolor="#800000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_binding    2197..2303
                     /locus_tag="attB region"
                     /label="attB region"
                     /ApEinfo_label="attB region"
                     /ApEinfo_fwdcolor="#6495ed"
                     /ApEinfo_revcolor="#6495ed"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     transit_peptide 3289..3345
                     /locus_tag="egl-13 NLS"
                     /label="egl-13 NLS"
                     /ApEinfo_label="egl-13 NLS"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2842..2892
                     /locus_tag="Synthetic Intron 3"
                     /label="Synthetic Intron 3"
                     /ApEinfo_label="Synthetic Intron 3"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2660..2710
                     /locus_tag="Synthetic Intron 2"
                     /label="Synthetic Intron 2"
                     /ApEinfo_label="Synthetic Intron 2"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2480..2530
                     /locus_tag="Synthetic Intron 1"
                     /label="Synthetic Intron 1"
                     /ApEinfo_label="Synthetic Intron 1"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     sig_peptide     2341..2373
                     /locus_tag="SV40 nls"
                     /label="SV40 nls"
                     /ApEinfo_label="SV40 nls"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    3352..3753
                     /locus_tag="myo-2 3' UTR"
                     /label="myo-2 3' UTR"
                     /ApEinfo_label="myo-2 3' UTR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    3118..3168
                     /locus_tag="Synthetic Intron 4"
                     /label="Synthetic Intron 4"
                     /ApEinfo_label="Synthetic Intron 4"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(5029..5390)
                     /locus_tag="Prps-0"
                     /label="Prps-0"
                     /ApEinfo_label="Prps-0"
                     /ApEinfo_fwdcolor="#339933"
                     /ApEinfo_revcolor="#339933"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1550..1583
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(2374..2479,2531..2659,2711..2841,2893..3117,
                     3169..3288)
                     /locus_tag="NeonGreen"
                     /label="NeonGreen"
                     /ApEinfo_label="NeonGreen"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     2304..2337
                     /locus_tag="FRT"
                     /label="FRT"
                     /ApEinfo_label="FRT"
                     /ApEinfo_fwdcolor="#800000"
                     /ApEinfo_revcolor="#800000"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(4233..5027)
                     /locus_tag="NeoR"
                     /label="NeoR"
                     /ApEinfo_label="NeoR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 1 offset 0"
     3'UTR           complement(3880..4231)
                     /locus_tag="unc-54 3'UTR"
                     /label="unc-54 3'UTR"
                     /ApEinfo_label="unc-54 3'UTR"
                     /ApEinfo_fwdcolor="#990000"
                     /ApEinfo_revcolor="#cc0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1708..1835
                     /locus_tag="MCS"
                     /label="MCS"
                     /ApEinfo_label="MCS"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2227..2228
                     /locus_tag="recombination site"
                     /label="recombination site"
                     /ApEinfo_label="recombination site"
                     /ApEinfo_fwdcolor="#ff9300"
                     /ApEinfo_revcolor="#ff9300"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
       61 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      121 tgctgcaatg ataccgcgag aTccacgctc accggctcca gatttatcag caataaacca
      181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      961 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     1021 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1081 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1141 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1201 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1261 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1321 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1381 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1441 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     1501 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaaA TAACTTCGTA
     1561 TAGCATACAT TATACGAAGT TATgttgggt aacgccaggg ttttcttgtg tggaattgtg
     1621 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1681 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctCTAGATGG
     1741 CGAAGAGCcc atggatccac tagtaacata tgtgcaggcc tGCTCTTCGG TAagcttatc
     1801 gataccgtcg acccgttaac cgggggcccg gtacccaatt cgccctatag tgagtcgtat
     1861 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact ggcatacgag ccggaagcat
     1921 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1981 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2041 cgcggggaga ggcggtttgc gtattgggcg aattccgctt cctcgctcac tgactcgctg
     2101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2161 tccacagaat caggggataa cgcaggaaag aacatgAAGC CGCGGTGCGG GTGCCAGGGC
     2221 GTGCCCTTGG GCTCCCCGGG CGCGTACTCC ACCTCACCCA TCTGGTCCAT CATGATGAAC
     2281 GGGTCGAGGT GGCGGTAGTT GATGAAGTTC CTATTCtcta gaaaGTATAG GAACTTCAAA
     2341 ATGGGATCCC CAAAGAAGAA GCGTAAGGTG TCGATGGTGT CGAAGGGAGA AGAGGATAAC
     2401 ATGGCTTCAC TCCCAGCTAC ACACGAACTC CACATCTTCG GATCGATCAA CGGAGTGGAT
     2461 TTCGATATGG TCGGACAAGg taagtttaaa catatatata ctaactaacc ctgattattt
     2521 aaattttcag GAACTGGAAA CCCAAACGAT GGATACGAGG AACTCAACCT CAAGTCGACA
     2581 AAGGGAGATC TGCAATTCTC GCCATGGATT CTCGTGCCAC ACATCGGATA CGGATTCCAC
     2641 CAATACCTCC CATACCCAGg taagtttaaa ctgagttcta ctaactaacg agtaatattt
     2701 aaattttcag ATGGAATGTC ACCATTCCAA GCTGCCATGG TGGATGGATC GGGATACCAA
     2761 GTTCACCGAA CAATGCAATT CGAGGATGGA GCCTCGCTCA CAGTGAACTA CCGATACACA
     2821 TACGAGGGAT CGCACATCAA Ggtaagttta aacagttcgg tactaactaa ccatacatat
     2881 ttaaattttc agGGAGAGGC TCAAGTTAAG GGAACAGGAT TCCCAGCTGA TGGACCAGTG
     2941 ATGACAAACT CACTCACAGC TGCTGATTGG TGCCGATCGA AAAAGACATA CCCAAATGAT
     3001 AAGACAATCA TCTCGACATT CAAGTGGTCG TACACTACTG GAAACGGAAA GCGATACCGA
     3061 TCGACAGCCC GAACAACATA CACATTCGCT AAGCCAATGG CCGCCAACTA CCTCAAGgta
     3121 agtttaaaca tgattttact aactaactaa tctgatttaa attttcagAA TCAACCAATG
     3181 TACGTGTTCC GAAAGACAGA ACTCAAGCAC TCAAAGACAG AGCTGAACTT CAAAGAGTGG
     3241 CAAAAGGCCT TCACAGATGT GATGGGAATG GATGAACTCT ACAAGGCCTC CCGTCGTCGT
     3301 AAGGCCAACC CAACCAAGCT CTCCGAGAAC GCCAAGAAGC TCGCCAAGta gatcaactgc
     3361 tctaattcca tctcgtttct ctcttttctt tcaatctctt tacccgtttc tcttccccta
     3421 cgtcccccaa acctaccact gcacagtaac acatacacat tcaacgtgtc acaataaaag
     3481 cccatcaaca ccttctctgt tgtttttttt cttcaaatgc atttttttca ctttgtttaa
     3541 taatttttac gcatcggatt tgtaggatag tggaaatttg tccttcaatg cttcaattga
     3601 ttcgacactt ctcaatattt ttgatcaata aaccattgat tttctattta gaagaagatc
     3661 actgtaaata acaccaaaaa catcacagca aaaaaactcc cactcaatta tattactgtt
     3721 tttattgaaa ctagaataaa tttgaaaatc ttcaaagcat acacaattaa aaaaaacgaa
     3781 aaaagaaaaa gtcccgttga ttctcaaact taaattttta aacttgtctg aaccatgttg
     3841 taagtttgta aaagtttccc agttgtcctc gaacaGGTCT AAAAATTGCG GTCATAAACT
     3901 GAAACGTAAC ATATGATAAG GTATTTTGTG TGCGGTTTTT TCTATGATGT TTCAAAAAAA
     3961 TTTATCAGAA GTAAAAAACA TAAGAAGTGT ACACAGAACA TTGTGTAGAT TTTGGGATAG
     4021 AATTATTATT AATGGGGGGA GGGAGCACAA TTTTTTTCGA CTTTTTATTA CGAATTAATT
     4081 CTATTTTTGA ATCTACACAA TTTCATTGTT AGAGGTGACT TAAAAGAAGC TAAAAAACAA
     4141 AGAAATTAAG AGAAGTTTTT TGATAATAAC AAAAATAGGG GGTGGGAGCA CAGGGAGAAA
     4201 GAGCATGTAG GGATGTTGAA GAGTAATTGG ACTCAGAAGA ACTCGTCAAG AAGGCGATAG
     4261 AAGGCGATGC GCTGCGAATC GGGAGCGGCG ATACCGTAAA GCACGAGGAA GCGGTCAGCC
     4321 CATTCGCCGC CAAGCTCCTC AGCAATATCA CGGGTAGCCA ACGCTATGTC CTGATAGCGG
     4381 TCCGCCACAC CCAGCCGGCC ACAGTCGATG AATCCAGAAA AGCGGCCATT TTCCACCATG
     4441 ATATTCGGCA AGCAGGCATC GCCATGGGTC ACGACGAGAT CCTCGCCGTC GGGCATGCGC
     4501 GCCTTGAGCC TGGCGAACAG TTCGGCTGGT GCGAGCCCCT GATGTTCTTC GTCCAGATCA
     4561 TCCTGATCGA CAAGACCGGC TTCCATCCGA GTACGTGCTC GCTCGATGCG ATGTTTCGCT
     4621 TGGTGGTCGA ATGGGCAGGT AGCCGGATCA AGCGTATGCA GCCGCCGCAT TGCATCAGCC
     4681 ATGATGGATA CTTTCTCGGC AGGAGCAAGG TGAGATGACA GGAGATCCTG CCCCGGCACT
     4741 TCGCCCAATA GCAGCCAGTC CCTTCCCGCT TCAGTGACAA CGTCGAGCAC AGCTGCGCAA
     4801 GGAACGCCCG TCGTGGCCAG CCACGATAGC CGCGCTGCCT CGTCCTGCAG TTCATTCAGG
     4861 GCACCGGACA GGTCGGTCTT GACAAAAAGA ACCGGGCGCC CCTGCGCTGA CAGCCGGAAC
     4921 ACGGCGGCAT CAGAGCAGCC GATTGTCTGT TGTGCCCAGT CATAGCCGAA TAGCCTCTCC
     4981 ACCCAAGCGG CCGGAGAACC TGCGTGCAAT CCATCTTGTT CAATCATATT ACCTTAAAAT
     5041 TCAAAAATTA ATTTCAGATC AGTCATAACC AAGAAAACAA AATAAACGTT AGGACAGAAA
     5101 ATTCAAATTA ATTAAAAATT ATGCTGAATA AGAAAATTAC ATGCAAAGCC TTATTTTCCT
     5161 CATAGAAACA GTTGAATAAC ATATTTTTTA ATTGAAAACA TAGCAAAATT CGTGGTAAAA
     5221 TTTGTCAAAA CCTTGGAAAA AATCGAAAAA AGAACATTTC CCGATGCATT TTCCTATTTC
     5281 TCGTTGCCGA CAACGTATAA ACGTTGAATA CTCAGTCAAC GAATTTGGCT GCTGGCTTGG
     5341 ACTCTCGCAC GGAAGAGACG CGTGTGTAGA TTTACGACGA AAGCAAAAAT gatcccgctt
     5401 tccgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5701 caagctgggc tgtgtgcacg aaccccccat tcagcccgac cgctgcgcct tatccggtaa
     5761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6181 atcaatctaa agtatatatg agtaaacttg gtctgacagt ta 
//
