LOCUS       pAP01                   4195 bp    DNA     circular SYN 09-MAY-2022
DEFINITION  sfGFP under Pbind with no EL222.
ACCESSION   JX560343
KEYWORDS    .
SOURCE      Cloning vector pSEVA531
  ORGANISM  Cloning vector pSEVA531
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 3460)
  AUTHORS   Silva-Rocha,R., Martinez-Garcia,E., Chavarria,M., Calles,B.,
            Arce-Rodriguez,A., de, las, Heras,A., Paez-Espino,D.,
            Durante-Rodriguez,G., Kim,J., Nikel,P.I., Platero,R., de and
            Lorenzo,V.
  TITLE     The Standard European Vector Architecture (SEVA): a coherent
            platform for analysis and deployment of complex prokaryotic
            phenotypes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3460)
  AUTHORS   Silva-Rocha,R., Martinez-Garcia,E., Chavarria,M., Calles,B.,
            Arce-Rodriguez,A., de, las, Heras,A., Paez-Espino,D.,
            Durante-Rodriguez,G., Kim,J., Nikel,P.I., Platero,R., de and
            Lorenzo,V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2012) Systems Biology Program, Centro Nacional de
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid,
            Madrid 28049, Spain
COMMENT     ##Assembly-Data-START## Assembly Method :: ApE v. v2.0.40
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4195
                     /organism="Cloning vector pSEVA531"
                     /mol_type="other DNA"
                     /db_xref="taxon:1246754"
     misc_feature    1..8
                     /Recognition_pattern="TTAAT^TAA"
                     /annotation_group="TTAATTAA"
                     /label="PacI"
                     /note="Geneious type: restriction site"
     primer_bind     9..>30
                     /note="R24"
     misc_feature    complement(10..30)
                     /created_by="primer3"
                     /annotation_group="1464728312013310000a2781707-4dc7-486e-8
                     6c9-02cef1787ee8"
                     /%GC="42.9"
                     /Tm="55.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="CTGTGTGAAATTGTTATCCGC"
                     /label="30 R"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     complement(10..30)
                     /Sequence="CTGTGTGAAATTGTTATCCGC"
                     /Hairpin_Tm="None"
                     /Tm="55.9"
                     /Self_Dimer_Tm="None"
                     /%GC="42.9"
                     /created_by="primer3"
                     /label="PSEVRI"
     misc_feature    31..34
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     motif           35..52
                     /created_by="User"
                     /modified_by="User"
                     /label="EL222 binding region"
     regulatory      35..87
                     /created_by="User"
                     /modified_by="User"
                     /label="EL222 binding site"
     motif           53..58
                     /created_by="User"
                     /label="-35"
     misc_feature    complement(61..87)
                     /created_by="primer3"
                     /annotation_group="82864832631847c7924f3-b4af-4569-bb6c-c5
                     3a602b5954"
                     /%GC="25.9"
                     /Tm="55.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TTTATTCGACTATAACAAACCATTTTC"
                     /label="83 R"
                     /note="Geneious type: primer_bind_reverse"
     motif           74..79
                     /created_by="User"
                     /label="-10"
     Insulator       88..102
                     /created_by="User"
                     /modified_by="User"
                     /label="Insulator "
     misc_feature    complement(88..102)
                     /created_by="primer3"
                     /annotation_group="1356074501976748d4d8bc11-3173-420c-8dc4
                     -ef890b711a0f"
                     /%GC="33.3"
                     /Tm="40.8"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="AATGAATTCTGTGTG"
                     /label="98 R"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    94..99
                     /Recognition_pattern="G^AATTC"
                     /annotation_group="GAATTC"
                     /label="EcoRI"
                     /note="Geneious type: restriction site"
     RBS             103..115
                     /created_by="User"
                     /label="RBSD"
     Insulator       116..121
                     /created_by="User"
     source          <122..>838
                     /organism="pSR59.4"
                     /mol_type="other DNA"
     CDS             122..838
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNG
                     IKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK*"
                     /modified_by="azadyp"
                     /label="sfgfp"
     primer_bind     122..140
                     /created_by="primer3"
                     /annotation_group="8680416279558fa244962-6897-4c5f-ad85-e7
                     4d47a9e08a"
                     /%GC="52.6"
                     /Tm="58.6"
                     /Hairpin_Tm="47.5"
                     /Self_Dimer_Tm="None"
                     /Sequence="ATGCGTAAAGGCGAAGAGC"
                     /label="118 F"
     misc_feature    283..288
                     /Recognition_pattern="GGTAC^C"
                     /annotation_group="GGTACC"
                     /label="KpnI"
                     /note="Geneious type: restriction site"
     misc_feature    302..331
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNG
                     IKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK*"
                     /label="Y66 (EGFP)"
     misc_feature    350..355
                     /Recognition_pattern="CA^TATG"
                     /annotation_group="CATATG"
                     /label="NdeI"
                     /note="Geneious type: restriction site"
     primer_bind     complement(626..642)
                     /Sequence="CCATCCTCCACGTTGTG"
                     /Hairpin_Tm="None"
                     /Tm="54.7"
                     /Self_Dimer_Tm="None"
                     /%GC="58.8"
                     /created_by="primer3"
                     /label="spiece1r"
     misc_feature    770..775
                     /Recognition_pattern="CA^TATG"
                     /annotation_group="CATATG"
                     /label="NdeI"
                     /note="Geneious type: restriction site"
     misc_feature    828..833
                     /Recognition_pattern="T^GTACA"
                     /annotation_group="TGTACA"
                     /label="BsrGI"
                     /note="Geneious type: restriction site"
     primer_bind     843..861
                     /created_by="primer3"
                     /annotation_group="1852208263494476543660a8-b56a-4978-bc21
                     -c5c332ead226"
                     /%GC="57.9"
                     /Tm="55.4"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="7.7"
                     /Sequence="GGGATCCTCTAGAGTCGAC"
                     /label="76 F"
     primer_bind     843..861
                     /Sequence="GGGATCCTCTAGAGTCGAC"
                     /Hairpin_Tm="None"
                     /Tm="55.4"
                     /Self_Dimer_Tm="7.7"
                     /%GC="57.9"
                     /created_by="primer3"
                     /label="PSEVFI"
     misc_feature    844..849
                     /Recognition_pattern="G^GATCC"
                     /annotation_group="GGATCC"
                     /label="BamHI"
                     /note="Geneious type: restriction site"
     misc_feature    850..855
                     /Recognition_pattern="T^CTAGA"
                     /annotation_group="TCTAGA"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     misc_feature    862..867
                     /Recognition_pattern="CTGCA^G"
                     /annotation_group="CTGCAG"
                     /label="PstI"
                     /note="Geneious type: restriction site"
     misc_feature    874..879
                     /Recognition_pattern="A^AGCTT"
                     /annotation_group="AAGCTT"
                     /label="HindIII"
                     /note="Geneious type: restriction site"
     misc_feature    880..887
                     /Recognition_pattern="GC^GGCCGC"
                     /annotation_group="GCGGCCGC"
                     /label="NotI"
                     /note="Geneious type: restriction site"
     primer_bind     complement(888..911)
                     /note="F24"
     primer_bind     897..911
                     /created_by="primer3"
                     /annotation_group="1464728234526319000659a1140-3f36-442c-9
                     943-d22dd699f388"
                     /%GC="66.7"
                     /Tm="54.0"
                     /Hairpin_Tm="34.5"
                     /Self_Dimer_Tm="None"
                     /Sequence="GGGAAAACCCTGGCG"
                     /label="130 F"
     misc_feature    912..917
                     /Recognition_pattern="A^CTAGT"
                     /annotation_group="ACTAGT"
                     /label="SpeI"
                     /note="Geneious type: restriction site"
     regulatory      918..1020
                     /regulatory_class="terminator"
                     /note="T0"
     misc_feature    1047..1054
                     /Recognition_pattern="ATTT^AAAT"
                     /annotation_group="ATTTAAAT"
                     /label="SwaI"
                     /note="Geneious type: restriction site"
     primer_bind     1053..1076
                     /Sequence="ATTTGACAGCTTATCATCGATAAA"
                     /Hairpin_Tm="34.6"
                     /Tm="54.8"
                     /Self_Dimer_Tm="1.2"
                     /%GC="29.2"
                     /created_by="primer3"
                     /label="spiece2f"
     misc_feature    complement(1076..1097)
                     /created_by="primer3"
                     /annotation_group="18524938426575953ffa7088-00b3-4f13-8e9e
                     -7b701b865986"
                     /%GC="36.4"
                     /Tm="55.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TGATAAACTACCGCATTACAGT"
                     /label="330 R"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1076..1097)
                     /created_by="User"
                     /Sequence="TGATAAACTACCGCATTACAGT"
                     /label="PSEVRII"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    1097^1098
                     /Original_Bases="CAGTTAAATTGCTAACGCAGTCAGGCACCGTG"
                     /label="CAGTTAAATTGCTAACGCAGTCAGGCACCGTG"
                     /note="Geneious type: Editing History Deletion"
     primer_bind     1098..1119
                     /created_by="primer3"
                     /annotation_group="185257830681239702423394-8d52-4bf1-8ac1
                     -fc8d1def6352"
                     /%GC="36.4"
                     /Tm="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="9.7"
                     /Sequence="TATGAAATCTAACAATGCGCTC"
                     /label="363 F"
     primer_bind     join(1098..1119,1098..1119)
                     /Sequence="TATGAAATCTAACAATGCGCTC"
                     /Hairpin_Tm="None"
                     /Tm="55.0"
                     /Self_Dimer_Tm="9.7"
                     /%GC="36.4"
                     /created_by="primer3"
                     /label="PSEVFII"
     primer_bind     complement(1119..1135)
                     /Sequence="TGCCGAGGATGACGATG"
                     /Hairpin_Tm="None"
                     /Tm="56.3"
                     /Self_Dimer_Tm="None"
                     /%GC="58.8"
                     /created_by="primer3"
                     /label="spiece2r"
     gene            1171..2289
                     /gene="tetA"
     CDS             1171..2289
                     /gene="tetA"
                     /codon_start=1
                     /transl_table=11
                     /product="tetracycline efflux transporter"
                     /protein_id="AFV59807.1"
                     /db_xref="GI:410073724"
                     /translation="MPVLPGLLRDIVHSDSIASHYGVLLALYALMQFLCAPVLGALSD
                     RFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRIVAGITGATGAVAGAYIADITDG
                     EDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCFLMQES
                     HKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVIFGEDRFR
                     WSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVLLAFATRG
                     WMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITGPLIVTAI
                     YAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    1242..1247
                     /Recognition_pattern="G^CTAGC"
                     /annotation_group="GCTAGC"
                     /label="NheI"
                     /note="Geneious type: restriction site"
     misc_feature    2290..2299
                     /Recognition_pattern="GACNN^NNGTC"
                     /annotation_group="GACNNNNGTC"
                     /label="PshAI"
                     /note="Geneious type: restriction site"
     misc_feature    2300..2545
                     /note="oriT"
     primer_bind     2526..2540
                     /created_by="primer3"
                     /annotation_group="1466438986464120000a739800f-3714-4d39-8
                     8dc-9174f0f746b6"
                     /%GC="73.3"
                     /Tm="56.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="1.8"
                     /Sequence="GCTCTGCGAGGCTGG"
                     /label="1,791 F"
     misc_feature    2546..2553
                     /Recognition_pattern="GGCCGG^CC"
                     /annotation_group="GGCCGGCC"
                     /label="FseI"
                     /note="Geneious type: restriction site"
     gene            complement(2554..3216)
                     /gene="rep"
     CDS             complement(2554..3216)
                     /gene="rep"
                     /codon_start=1
                     /transl_table=11
                     /product="replication protein"
                     /protein_id="AFV59808.1"
                     /db_xref="GI:410073725"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLL
                     HHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAY
                     VVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGP
                     GEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     primer_bind     2554..2566
                     /created_by="primer3"
                     /annotation_group="1464795714538271000c0396fd5-97a1-4f32-b
                     9af-739427d4aec3"
                     /%GC="84.6"
                     /Tm="56.4"
                     /Hairpin_Tm="53.1"
                     /Self_Dimer_Tm="24.5"
                     /Sequence="CTACCGGCGCGGC"
                     /label="1,819 F"
     misc_feature    complement(4055..4070)
                     /created_by="primer3"
                     /annotation_group="14664392394251400001a54adc0-9179-42f5-a
                     e63-c982159983f3"
                     /%GC="62.5"
                     /Tm="55.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Sequence="TACGGGCTTGCTCTCC"
                     /label="3,335 R"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    4076..4083
                     /Recognition_pattern="GG^CGCGCC"
                     /annotation_group="GGCGCGCC"
                     /label="AscI"
                     /note="Geneious type: restriction site"
     regulatory      4090..4194
                     /regulatory_class="terminator"
                     /note="T1"
     primer_bind     4149..4166
                     /Sequence="CGAAAGGCCCAGTCTTTC"
                     /Hairpin_Tm="39.0"
                     /Tm="55.7"
                     /Self_Dimer_Tm="10.9"
                     /%GC="55.6"
                     /created_by="primer3"
                     /label="spiece1f"
ORIGIN      
        1 ttaattaaag cggataacaa tttcacacag agccggtagc ctttagtcca tgttagcgaa
       61 gaaaatggtt tgttatagtc gaataaacac acagaattca ttaagaagga gatatggtac
      121 catgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
      181 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
      241 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct
      301 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
      361 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
      421 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
      481 aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa
      541 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
      601 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
      661 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
      721 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
      781 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgaac
      841 acgggatcct ctagagtcga cctgcaggca tgcaagcttg cggccgcgtc gtgactggga
      901 aaaccctggc gactagtctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      961 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag
     1021 gggtccccaa taattacgat ttacgtattt aaatttgaca gcttatcatc gataaactgt
     1081 aatgcggtag tttatcatat gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc
     1141 accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct cttgcgggat
     1201 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg
     1261 atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg ccgccgccca
     1321 gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc gaccacaccc
     1381 gtcctgtgga ttctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     1441 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     1501 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     1561 ggcgccatct ccctgcacgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
     1621 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgccgtcc gatgcccttg
     1681 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgaccat tgtggccgca
     1741 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     1801 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     1861 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     1921 ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg
     1981 gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc
     2041 atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga
     2101 cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcactgg accgctgatc
     2161 gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc
     2221 gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     2281 tcgacctgag acaattgtcc ttttccgctg cataaccctg cttcggggtc attatagcga
     2341 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     2401 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     2461 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     2521 atcctgctct gcgaggctgg ccgtaggccg gccctaccgg cgcggcagcg ttacccgtgt
     2581 cggcggctcc aacggctcgc catcgtccag aaaacacggc tcatcgggca tcggcaggcg
     2641 ctgctgcccg cgccgttccc attcctccgt ttcggtcaag gctggcaggt ctggttccat
     2701 gcccggaatg ccgggctggc tgggcggctc ctcgccgggg ccggtcggta gttgctgctc
     2761 gcccggatac agggtcggga tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg
     2821 gtcgtcgtga tcaaccacca cggcggcact gaacaccgac aggcgcaact ggtcgcgggg
     2881 ctggccccac gccacgcggt cattgaccac gtaggccgac acggtgccgg ggccgttgag
     2941 cttcacgacg gagatccagc gctcggccac caagtccttg actgcgtatt ggaccgtccg
     3001 caaagaacgt ccgatgagct tggaaagtgt cttctggctg accaccacgg cgttctggtg
     3061 gcccatctgc gccacgaggt gatgcagcag cattgccgcc gtgggtttcc tcgcaataag
     3121 cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc cagtgaccgg gcttgttctt
     3181 ggcttgaatg ccgatttctc tggactgcgt ggccatgctt atctccatgc ggtaggggtg
     3241 ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt
     3301 atgcgttgcg taaaagtggc agtcaattac agattttctt taacctacgc aatgagctat
     3361 tgcggggggt gccgcaatga gctgttgcgt accccccttt tttaagttgt tgatttttaa
     3421 gtctttcgca tttcgcccta tatctagttc tttggtgccc aaagaagggc acccctgcgg
     3481 ggttccccca cgccttcggc gcggctcccc ctccggcaaa aagtggcccc tccggggctt
     3541 gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc gctgccccct tggaaccccc
     3601 gcactcgccg ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc gactgccccc
     3661 actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc
     3721 gcgagccttg acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca
     3781 ggccggaggc ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc
     3841 gcagttggag ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg
     3901 tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc
     3961 cgagcgaagc gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg
     4021 caagggagcg cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggggggcgc
     4081 gcccagctgt ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     4141 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcct
//
