LOCUS       pATF51_AT_1step_pCOMB3X 4356 bp ds-DNA     circular     26-AUG-2025
DEFINITION  .
FEATURES             Location/Qualifiers
     rep_origin      1..683
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          1016..1036
                     /label="OATC47"
                     /note="sequence: gttgtgtggaattgtgagcgg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_binding    1023..1045
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             1066..1131
                     /label="Translation 1066-1131"
                     /translation="MKKTAIAIAVALAGFATVAQAA"
     CDS             1066..1131
                     /label="Translation 1066-1131"
                     /translation="MKKTAIAIAVALAGFATVAQAA"
     misc_feature    1083..1102
                     /label="ompA FOR primer"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_fwdcolor="#ff0080"
     misc_feature    1119..1131
                     /label="SfiI site at 5' end of L chain"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          1119..1152
                     /label="OATF154"
                     /note="sequence: ggcccaggctgccaaGTCTTCatggtgagcaagg"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(1119..1152)
                     /label="OATF155"
                     /note="sequence: ccttgctcaccatGAAGACttggcagcctgggcc"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          1121..1153
                     /label="OATF158"
                     /note="sequence: cccaggctgccaaGTCTTCatggtgagcaaggg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(1121..1153)
                     /label="OATF159"
                     /note="sequence: cccttgctcaccatGAAGACttggcagcctggg"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(1122..1145)
                     /label="OATC68"
                     /note="sequence: ACTGGTGAAGACTTGGCAGCCTGG"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             1140..1142
                     /label="Translation 1140-1142"
                     /translation="M"
     misc_feature    1140..1859
                     /label="GFP"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             1140..1859
                     /label="Translation 1140-1859"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGCYKTRAEVKFEGDTLVNRIELHGIDFKEDGNILGHKLEYNFNSHNVYIMPDKQNNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDYHYLHTWSELSKDPNEKRDHMVLLEFVEAAGITLGMDELYK*"
     primer          complement(1402..1419)
                     /label="OATF47"
                     /note="sequence: cgtagccttcgggcatgg"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          1844..1880
                     /label="OATF160"
                     /note="sequence: cgagctgtacaagtaaGAAGACccgctAGCaccaagg"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(1844..1880)
                     /label="OATF161"
                     /note="sequence: ccttggtGCTagcggGTCTTCttacttgtacagctcg"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          1847..1881
                     /label="OATF156"
                     /note="sequence: gctgtacaagtaaGAAGACccgctAGCaccaaggg"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(1847..1881)
                     /label="OATF157"
                     /note="sequence: cccttggtGCTagcggGTCTTCttacttgtacagc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          1854..1876
                     /label="OATC67"
                     /note="sequence: GAGATAGAAGACCCGCTAGCACC"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1868..1873
                     /label="NheI site between VH and CH1"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    1868..2187
                     /label="Fd domain"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             1868..2187
                     /label="Translation 1868-2187"
                     /translation="ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT"
     misc_feature    1874..2187
                     /label="Fd domain - CH1"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          1879..1902
                     /label="OATF146"
                     /note="sequence: gggcccatcggtgTTCCCCCTGGC"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(1879..1902)
                     /label="OATC147"
                     /note="sequence: GCCAGGGGGAAcaccgatgggccc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(2131..2150)
                     /label="OATF13"
                     /note="sequence: CCACCTTGGTGTTGCTGGGC"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    2183..2203
                     /label="alreadythere"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             2183..2317
                     /label="Translation 2183-2317"
                     /translation="TSPDYASGGSHHHHHH*EGGGSEGGGSEGGGSEGGGSGGGSGSGD"
     primer          2184..2247
                     /label="OATF66"
                     /note="sequence: CTAGTCCGGActacgcttctggcggatcccatcaccatcaccatcactagGAGGGTGGTGGCTC"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(2184..2247)
                     /label="OATF67"
                     /note="sequence: GAGCCACCACCCTCctagtgatggtgatggtgatgggatccgccagaagcgtagTCCGGACTAG"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    2188..2193
                     /label="BspEI 3' end cloning site for Fd domain"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    2194..2230
                     /label="GSH6 tag"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2231..2233
                     /label="Amber stop codon after 3' end of Fddomain"
                     /ApEinfo_revcolor="#f420ff"
                     /ApEinfo_fwdcolor="#f420ff"
     misc_feature    2234..2767
                     /label="C-terminal half of M13 P3"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             2234..2767
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="EGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTENADENVLQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMYVFSTFANILRNKES*"
     misc_feature    2433..2457
                     /label="p3 REV/FOR primers"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_fwdcolor="#ff0080"
     misc_feature    2749..2767
                     /label="NheI remover primer area 3'"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2768..2788
                     /label="NheI remover primer area 5'"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2781..2788
                     /label="NotI site at 3' end of M13 P3"
                     /ApEinfo_revcolor="#ff081a"
                     /ApEinfo_fwdcolor="#ff081a"
     primer          complement(2937..2960)
                     /label="OATF40"
                     /note="sequence: ccaaactggaacaacactcaaccc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     rep_origin      complement(2947..3253)
                     /label="F1 ori"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             3600..4259
                     /label="AmpR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             3600..4259
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGTRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggcagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      901 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      961 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1021 gtggaattgt gagcggataa caattgaatt caggaggaat ttaaaatgaa aaagacagct
     1081 atcgcgattg cagtggcact ggctggtttc gctaccgtgg cccaggctgc caaGTCTTCa
     1141 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1201 gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat gccacctacg
     1261 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1321 tcgtgaccac cctgacctac ggcgtccagt gcttcagccg ctaccccgac cacatgaagc
     1381 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1441 tcaaggacga cggctgctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1501 tgaaccgcat cgagctgcac ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1561 agctggagta caacttcaac agccacaacg tctatatcat gcccgacaag cagaacaacg
     1621 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1681 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gactaccact
     1741 acctgcacac ctggtccgag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1801 tgctggagtt cgtggaggcc gccgggatca ctctcggcat ggacgagctg tacaagtaaG
     1861 AAGACccgct AGCaccaagg gcccatcggt gTTCCCCCTG GCACCCTCCT CCAAGAGCAC
     1921 CTCTGGGGGC ACAGCGGCCC TGGGCTGCCT GGTCAAGGAC TACTTCCCCG AACCGGTGAC
     1981 GGTGTCGTGG AACTCAGGCG CCCTGACCAG CGGCGTGCAC ACCTTCCCGG CTGTCCTACA
     2041 GTCCTCAGGA CTCTACTCCC TCAGCAGCGT GGTGACCGTG CCCTCCAGCA GCTTGGGCAC
     2101 CCAGACCTAC ATCTGCAACG TGAATCACAA GCCCAGCAAC ACCAAGGTGG ACAAGAAAGT
     2161 TGAGCCCAAA TCTTGTGACA AAACTAGTCC GGActacgct tctggcggat cccatcacca
     2221 tcaccatcac tagGAGGGTG GTGGCTCTGA GGGTGGCGGT TCTGAGGGTG GCGGCTCTGA
     2281 GGGAGGCGGT TCCGGTGGTG GCTCTGGTTC CGGTGATTTT GATTATGAAA AGATGGCAAA
     2341 CGCTAATAAG GGGGCTATGA CCGAAAATGC CGATGAAAAC GTGCTACAGT CTGACGCTAA
     2401 AGGCAAACTT GATTCTGTCG CTACTGATTA CGGTGCTGCT ATCGATGGTT TCATTGGTGA
     2461 CGTTTCCGGC CTTGCTAATG GTAATGGTGC TACTGGTGAT TTTGCTGGCT CTAATTCCCA
     2521 AATGGCTCAA GTCGGTGACG GTGATAATTC ACCTTTAATG AATAATTTCC GTCAATATTT
     2581 ACCTTCCCTC CCTCAATCGG TTGAATGTCG CCCTTTTGTC TTTGGCGCTG GTAAACCATA
     2641 TGAATTTTCT ATTGATTGTG ACAAAATAAA CTTATTCCGT GGTGTCTTTG CGTTTCTTTT
     2701 ATATGTTGCC ACCTTTATGT ATGTATTTTC TACGTTTGCT AACATACTGC GTAATAAGGA
     2761 GTCTTAAtaa ttaatttaaG CGGCCGCAga tctGCTCTCT GAGGAGGATC Tgggaaattg
     2821 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2881 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2941 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3001 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3061 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3121 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3181 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3241 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3781 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3961 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4081 ggtttattgc tgataaatct ggagccggtg agcgtgggac tcgcggtatc attgcagcac
     4141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4321 ttaaaaggat ctaggtgaag atcctttttg ataatc
//