LOCUS       Exported                3962 bp ds-DNA     circular SYN 25-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3962)
  AUTHORS   Matt
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 9, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3962
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          74..868
                     /organism="Escherichia coli str. K-12 substr. MG1655"
                     /strain="K-12"
                     /sub_strain="MG1655"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:511145"
     source          869..1123
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          869..1123
                     /organism="Escherichia species"
                     /strain="strain"
                     /mol_type="genomic DNA"
     source          869..1123
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(1..36)
                     /label=MBA645
     promoter        1..35
                     /label=BBa_J23108
     primer_bind     complement(7..36)
                     /label=MBA695
     primer_bind     complement(11..35)
                     /label=JL064
     primer_bind     73..107
                     /label=MBA704
     CDS             74..868
                     /codon_start=1
                     /transl_table=11
                     /gene="thyA"
                     /gene_synonym="ECK2823;JW2795"
                     /locus_tag="b2827"
                     /product="thymidylate synthetase"
                     /function="enzyme; 2'-Deoxyribonucleotide metabolism"
                     /EC_number="2.1.1.45"
                     /label=thyA
                     /note="GO_component: GO:0005737 - cytoplasm; GO_process: 
                     GO:0015949 - nucleobase, nucleoside and nucleotide 
                     interconversion"
                     /db_xref="ASAP:ABE-0009268"
                     /db_xref="UniProtKB/Swiss-Prot:P0A884"
                     /db_xref="EcoGene:EG11002"
                     /experiment="N-terminus verified by Edman degradation: PMID
                     6308660"
                     /protein_id="AAC75866.1"
                     /translation="MKQYLELMQKVLDEGTQKNDRTGTGTLSIFGHQMRFNLQDGFPLV
                     TTKRCHLRSIIHELLWFLQGDTNIAYLHENNVTIWDEWADENGDLGPVYGKQWRAWPTP
                     DGRHIDQITTVLNQLKNDPDSRRIIVSAWNVGELDKMALAPCHAFFQFYVADGKLSCQL
                     YQRSCDVFLGLPFNIASYALLVHMMAQQCDLEVGDFVWTGGDTHLYSNHMDQTHLQLSR
                     EPRPLPKLIIKRKPESIFDYRFEDFEIEGYDPHPGIKAPVAI"
     gene            74..868
                     /gene="thyA"
                     /gene_synonym="ECK2823;JW2795"
                     /locus_tag="b2827"
                     /label=thyA
                     /db_xref="EcoGene:EG11002"
     primer_bind     complement(843..868)
                     /label=MBA703
     RBS             869..904
                     /label=RBS (4427)
     primer_bind     869..894
                     /label=AGR822
     CDS             905..1123
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="ODNLECBM_00895"
                     /product="translation initiation factor IF-1 [Escherichia 
                     coli Nissle 1917]"
                     /label=infA
                     /inference="ab initio prediction:same species:2.6"
                     /inference="similar to AA sequence:same species:AID81871.1"
                     /translation="MAKEDNIEMQGTVLETLPNTMFRVELENGHVVTAHISGKMRKNYI
                     RILTGDKVTVELTPYDLSKGRIVFRSR"
     primer_bind     complement(1112..1123)
                     /label=AGR821
     primer_bind     1123..1156
                     /label=MBA696
     misc_feature    1124..1127
                     /label=BsaI Overlap
     primer_bind     1128..1157
                     /label=MBA286
     primer_bind     1128..1153
                     /label=AGR341
     primer_bind     complement(1155..1175)
                     /label=MBA489
     misc_feature    1182..1198
                     /label=113R
     primer_bind     1200..1220
                     /label=AGR526
     primer_bind     1200..1218
                     /label=MBA214
     primer_bind     complement(1201..1221)
                     /label=AGR366
     primer_bind     complement(1204..1225)
                     /label=MBA347
     primer_bind     complement(1207..1228)
                     /label=MBA015
     primer_bind     complement(1237..1262)
                     /label=AGR360
     primer_bind     complement(1257..1284)
                     /label=MBA177
     primer_bind     1281..1308
                     /label=AGR431
     primer_bind     complement(1322..1345)
                     /label=AGR301
     rep_origin      complement(1333..2283)
                     /direction=LEFT
                     /label=pSC101 ori
     primer_bind     1333..1360
                     /label=MBA206
     primer_bind     1991..2013
                     /label=MBA130
     primer_bind     complement(1991..2013)
                     /label=MBA129
     primer_bind     2212..2237
                     /label=JL025
     primer_bind     complement(2212..2237)
                     /label=AGR434
     RBS             2287..2299
                     /label=RBS for repA
     primer_bind     complement(2288..2323)
                     /label=MBA205
     promoter        complement(2304..2323)
                     /label=repA promoter
     primer_bind     2324..2354
                     /label=MBA228
     primer_bind     complement(2405..2431)
                     /label=MBA213
     primer_bind     complement(2665..2684)
                     /label=JL015
     promoter        2685..2771
                     /label=predicted promoter
     CDS             2807..3622
                     /codon_start=1
                     /label=kan-r
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    2837
                     /label=XhoI might be mutated
     primer_bind     2890..2909
                     /label=MBA714
     primer_bind     complement(2890..2909)
                     /label=MBA711
     primer_bind     3589..3611
                     /label=AGR037
     primer_bind     3664..3682
                     /label=MBA493
     primer_bind     3786..3809
                     /label=MBA567
     misc_feature    3795..3817
                     /label=pTET For
     primer_bind     3804..3826
                     /label=MBA016
     primer_bind     complement(3804..3818)
                     /label=MBA147
     terminator      3818..3946
                     /label=dbI term
     primer_bind     complement(3821..3843)
                     /label=AGR157
     primer_bind     3893..3919
                     /label=MBA215
     primer_bind     complement(3928..3962)
                     /label=MAB553
     primer_bind     complement(3929..3959)
                     /label=AGR295
     primer_bind     complement(3929..3957)
                     /label=AGR420
     primer_bind     join(3960..3962,1..35)
                     /label=MBA646
ORIGIN
        1 ctgacagcta gctcagtcct aggtataatg ctagctcgac cgctatttta gcccatatca
       61 gaggtaacac ctaatgaaac agtatttaga actgatgcaa aaagtgctcg acgaaggcac
      121 acagaaaaac gaccgtaccg gaaccggaac gctttccatt tttggtcatc agatgcgttt
      181 taacctgcaa gatggattcc cgctggtgac aactaaacgt tgccacctgc gttccatcat
      241 ccatgaactg ctgtggtttc tgcagggcga cactaacatt gcttatctac acgaaaacaa
      301 tgtcaccatc tgggacgaat gggccgatga aaacggcgac ctcgggccag tgtatggtaa
      361 acagtggcgc gcctggccaa cgccagatgg tcgtcatatt gaccagatca ctacggtact
      421 gaaccagctg aaaaacgacc cggattcgcg ccgcattatt gtttcagcgt ggaacgtagg
      481 cgaactggat aaaatggcgc tggcaccgtg ccatgcattc ttccagttct atgtggcaga
      541 cggcaaactc tcttgccagc tttatcagcg ctcctgtgac gtcttcctcg gcctgccgtt
      601 caacattgcc agctacgcgt tattggtgca tatgatggcg cagcagtgcg atctggaagt
      661 gggtgatttt gtctggaccg gtggcgacac gcatctgtac agcaaccata tggatcaaac
      721 tcatctgcaa ttaagccgcg aaccgcgtcc gctgccgaag ttgattatca aacgtaaacc
      781 cgaatccatc ttcgactacc gtttcgaaga ctttgagatt gaaggctacg atccgcatcc
      841 gggcattaaa gcgccggtgg ctatctaagg ttcaaattac ggtagtgata ccccagagga
      901 ttagatggcc aaagaagaca atattgaaat gcaaggtacc gttcttgaaa cgttgcctaa
      961 taccatgttc cgcgtagagt tagaaaacgg tcacgtggtt actgcacaca tctccggtaa
     1021 aatgcgcaaa aactacatcc gcatcctgac gggcgacaaa gtgactgttg aactgacccc
     1081 gtacgacctg agcaaaggcc gcattgtctt ccgtagtcgc tgaagcccga gaaaagatta
     1141 cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1201 cgagcggtat cagctcactc aaaggcggta atcccgggca gttttccctt tgatatctaa
     1261 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     1321 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     1381 tttttgcgtg agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa
     1441 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     1501 tcttagtccg ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     1561 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     1621 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     1681 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     1741 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     1801 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     1861 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     1921 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     1981 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     2041 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     2101 gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt
     2161 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     2221 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     2281 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     2341 atgataatta catgtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     2401 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     2461 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     2521 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     2581 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     2641 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat gttcgattta ttcaacaaag
     2701 ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2761 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2821 acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2881 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     2941 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3001 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3061 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     3121 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3181 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3241 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3301 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct
     3361 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3421 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3481 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3541 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3601 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3661 tacgctgact tgacgggact cgagattacc attattgaag catttatcag ggttattgtc
     3721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3781 catttcgaat tcgtgctagt gtagatcgct actagagcca ggcatcaaat aaaacgaaag
     3841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     3901 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatacaat tgtctagata
     3961 cc
//
