LOCUS       Exported                5013 bp ds-DNA     circular SYN 27-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Apr 25, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
COMMENT     Ap-TbpA gene
FEATURES             Location/Qualifiers
     source          1..5013
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    12..467
                     /label=f1 Ori
     CDS             599..1459
                     /codon_start=1
                     /label=bla
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    1663..2138
                     /label=OriV - ColE1/pBR322
     misc_feature    3348..3367
                     /label=oligo #24,  T7 promoter
                     /note="Tm 47.7 C"
     misc_feature    3414..3420
                     /label=rbs
                     /note="Ribosomal binding site (Shine-Dalgarno sequence) 
                     positioned appropriately in front of the start codon (ATG) 
                     within the Nde site to result in effective expression of 
                     coding sequences starting at the Nde1 site."
     CDS             3428..3487
                     /codon_start=1
                     /label=pelB signal peptide
                     /translation="MKYLLPTAAAGLLLLAAQPA"
     misc_feature    3487..3561
                     /label=oligo  4582
     CDS             3500..3541
                     /codon_start=1
                     /label=polyhisTEV
                     /translation="HHHHHHHHENLYFQ"
     misc_feature    3524..3541
                     /label=Untitled
     CDS             3542..3673
                     /codon_start=1
                     /label=SBP - streptavidin binding peptide
                     /translation="EFMDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREPSSGR"
     CDS             3674..3691
                     /codon_start=1
                     /label=TEV cleavage site
                     /translation="ENLYFQ"
     misc_feature    3692..3715
                     /label=New oligo
     misc_feature    3693..4845
                     /label=e4849*, pET2-His-TEV-SBP-TEV-KanR
                     /note="Ap-TbpA gene"
     misc_feature    3703..3722
                     /label=oligo 1014
     misc_feature    3736..3741
                     /label=-35_signal
                     /note="     -35_signal      94..99
                                          /label=-35
                                          /SECDrawAs=""Label""
                                          /SECName=""-35"""
     misc_feature    3756..3764
                     /label=-10_signal
                     /note="     -10_signal      114..122
                                          /label=-10
                                          /SECDrawAs=""Label""
                                          /SECName=""-10"""
     misc_feature    3812..3826
                     /label=oligo 532
     CDS             3815..4630
                     /codon_start=1
                     /label=KanR
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    3815..3831
                     /label=oligo 148
     misc_feature    3828..3847
                     /label=OLIGO 019 (Tn903A)
     misc_feature    complement(4510..4529)
                     /label=OLIGO 020 (TN903B)
     misc_feature    complement(4609..4634)
                     /label=oligo 983
     misc_feature    complement(4619..4636)
                     /label=oligo 149
     misc_feature    complement(4622..4632)
                     /label=oligo 533
     misc_feature    complement(4822..4850)
                     
     misc_feature    4840..4845
                     /label=e58 [168-1309]
                     /note="e058.e1834. pUK4K plasmid. The plasmid contains a 
                     kanR gene block from Tn903 inserted between MCS within lacZ
                     gene (N-terminal region)."
     CDS             4857..4874
                     /codon_start=1
                     /label=His6
                     /translation="HHHHHH"
     misc_feature    4942..4988
                     /label=T7 terminator
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     3361 tatagggaga ccacaacggt ttccctctag taataatttt gtttaacttt aagaaggaga
     3421 tatacatatg aaatacctgc tgccgaccgc tgctgctggt ctgctgctcc tcgctgccca
     3481 gccggcgatg gccatggccc atcatcatca tcaccatcac catgaaaacc tgtattttca
     3541 ggaattcatg gacgaaaaaa ccaccggttg gcgtggtggt cacgttgttg aaggtctggc
     3601 tggtgaactg gaacagctgc gtgctcgtct ggaacaccac ccgcagggtc agcgtgaacc
     3661 gagcagcggc agagaaaacc tgtattttca gggatccggg ggggaaagcc acgttgtgtc
     3721 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     3781 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     3841 ttgctcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3901 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     3961 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     4021 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4081 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
     4141 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4201 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4261 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4321 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4381 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4441 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4501 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4561 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     4621 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     4681 acgggacggc ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg
     4741 catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg
     4801 tccacctaca acaaagctct catcaaccgt ggctccctct ctagaagctt ctcgagcacc
     4861 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     4921 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     4981 gttttttgct gaaaggagga actatatccg gat
//
