LOCUS       Exported                6531 bp ds-DNA     circular SYN 18-DEC-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6531)
  AUTHORS   Kyle Watters
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 18, 2019 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6531
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(157..185)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
                     /note="This reverse directional feature has 3 segments:
                        1: 157 .. 162 / #ffffff / -10
                        2: 163 .. 179 / #ffffff
                        3: 180 .. 185 / #ffffff / -35"
     promoter        298..402
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             403..1263
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1:  403 ..  471 / #ccffcc / signal sequence
                        2:  472 .. 1263 / #ccffcc
                     Cleavage site after base 471"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1434..2022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    2208..2350
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     CDS             complement(2452..2643)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4204..4222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     RBS             4311..4333
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     CDS             4353..4382
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHHHHHH"
     CDS             4392..5492
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /note="color: #cc99b2"
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             5547..5567
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="color: #cc99b2
                     Cleavage site after base 5564"
                     /translation="ENLYFQS"
     misc_feature    5574..6137
                     /label=Prophage Region
                     /note="color: #a6acb3"
     CDS             5574..6137
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="NP71_RS07005"
                     /old_locus_tag="NP71_07010"
                     /product="hypothetical protein"
                     /label=AcrIIA13
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Protein Homology."
                     /note="color: #ff9900"
                     /inference="COORDINATES:same species:RefSeq"
                     /protein_id="WP_050337628.1"
                     /translation="MRETIQRLLASNLSSNSIALATGVSQAVISKLRNAKKEIGNLSLN
                     SAEKLYNYQKGLEVMNKSIEIKDQNNIVLIDSLGQFFTDIENDNNGRYNIDYVLLNEVE
                     HDNGNTYYEVGMYRTEEVPFSDKVTQDNVELLEDKWLQIDQQGESYVESIFFENEEDAR
                     EYIKLVLKGHETFEETAKAIGVIK"
     gene            5742..6137
                     /locus_tag="NP71_RS07005"
                     /old_locus_tag="NP71_07010"
                     /label=NP71_RS07005
                     /note="color: #a6acb3; direction: RIGHT"
     terminator      6275..6322
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
ORIGIN
        1 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
       61 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
      121 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
      181 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
      241 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
      301 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      361 taaccctgat aaatgcttca ataacattga aaaaggaaga gtatgagtat tcaacatttc
      421 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      481 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      541 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      601 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
      661 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      721 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      781 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      841 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      901 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
      961 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1021 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1081 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1141 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1201 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1261 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1321 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1381 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1441 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1501 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1561 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1621 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1681 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1741 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1801 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1861 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1921 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1981 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2041 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2101 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2161 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     2221 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     2281 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     2341 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2401 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2461 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     2521 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     2581 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2641 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     2701 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     2761 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     2821 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2881 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     2941 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     3001 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     3061 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     3121 aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     3181 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     3241 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     3301 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     3361 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     3421 ccagtgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     3481 tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa
     3541 gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg
     3601 tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3661 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3721 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3781 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     3841 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3901 ggtcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     3961 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4021 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4081 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4141 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4201 aattaatacg actcactata gggagaccac aacggtttcc ctctagtgcc ggctccggag
     4261 agctctttaa ttaagcggcc gccctgcagg actcgagttc tagaaataat tttgtttaac
     4321 tttaagaagg agatatacat atgaaatctt ctcaccatca ccatcaccat caccatcacc
     4381 atggttcttc tatgaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag
     4441 gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca
     4501 ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg
     4561 gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt
     4621 tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg
     4681 ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga
     4741 tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg
     4801 ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact
     4861 tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt
     4921 acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg
     4981 ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg
     5041 cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg
     5101 acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca
     5161 aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg
     5221 caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag
     5281 acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg aaagatccac
     5341 gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga
     5401 tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga
     5461 ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac aacaacaata
     5521 acaataacaa caacctcggg atcgaggaaa acctgtactt ccaatccaat gcaatgagag
     5581 agacaataca acgattgtta gcaagtaact tatccagcaa cagtattgca ttagcgactg
     5641 gagtatcgca agcagtgatt agtaagttgc gtaatgccaa gaaagaaata ggaaatctaa
     5701 gtttgaacag cgctgaaaaa ttatataatt atcaaaaggg gttggaagta atgaataaat
     5761 caatcgaaat taaagaccaa aataacatag tattaattga cagtttagga caattcttta
     5821 ctgacattga gaacgacaat aacggtcgtt acaacattga ctatgttttg ttgaatgaag
     5881 tggaacatga caatggcaac acgtattatg aagtgggaat gtacagaacc gaagaggttc
     5941 ctttcagtga taaagttact caagataatg ttgaattgct agaagataaa tggcttcaaa
     6001 tagatcaaca aggagaaagt tatgttgaaa gtatcttttt cgaaaacgaa gaagatgcga
     6061 gagagtatat caaattagtt ctgaaaggtc acgaaacttt tgaagaaact gcaaaagcaa
     6121 tcggcgtgat taaataaata ttggaagtgg ataacggatc cgcgatcgcg gcgcgccacc
     6181 tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga
     6241 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     6301 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg
     6361 gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg
     6421 ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca
     6481 acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa t
//
