LOCUS       Exported                7173 bp ds-DNA     circular SYN 06-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7173)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 29, 2024 from SnapGene Viewer 5.2.5.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7173
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_recomb     1..100
                     /locus_tag="RP4 oriT/RSK oriV"
                     /label=RP4 oriT/RSK oriV
                     /note="RP4 oriT/RSK oriV"
     misc_feature    1..20
                     /locus_tag="NBU1-f1v1"
                     /label=NBU1-f1v1
     misc_feature    115..118
                     /locus_tag="Part 1 overhang"
                     /label=Part 1 overhang
     misc_feature    120..125
                     /locus_tag="BsmBI site(1)"
                     /label=BsmBI site
                     /label=BsmBI site(1)
     promoter        138..166
                     /locus_tag="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    174..190
                     /locus_tag="lac operator"
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             210..533
                     /codon_start=1
                     /gene="lacZ fragment"
                     /locus_tag="lacZ-alpha"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     complement(509..528)
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectablemarker"
     misc_feature    546..551
                     /locus_tag="BsmBI site"
                     /label=BsmBI site
     misc_feature    553..556
                     /locus_tag="Part 7 overhang"
                     /label=Part 7 overhang
     misc_feature    557..579
                     /locus_tag="NBU1-attNseqFwd"
                     /label=NBU1-attNseqFwd
     misc_feature    659
                     /locus_tag="New Feature"
                     /label=New Feature
     misc_feature    681
                     /locus_tag="New Feature(1)"
                     /label=New Feature
                     /label=New Feature(1)
     CDS             complement(769..2106)
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="IntN1"
                     /product="IntN1"
                     /label=IntN1
                     /note="NBU1 integrase"
                     /protein_id="QJT41633.1"
                     /translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
                     LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
                     PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
                     GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
                     RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
                     FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
                     YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
                     IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
     misc_feature    775
                     /locus_tag="mut to C"
                     /label=mut to C
     misc_feature    777
                     /locus_tag="mut to G"
                     /label=mut to G
     misc_feature    980
                     /locus_tag="mut to T (silent)"
                     /label=mut to T (silent)
     misc_recomb     complement(2122..2451)
                     /locus_tag="attN1 region"
                     /label=attN1 region
                     /note="attN1 region"
     misc_feature    complement(2200..2220)
                     /locus_tag="NBU1-intSeqFwd"
                     /label=NBU1-intSeqFwd
     misc_feature    complement(2311..2324)
                     /locus_tag="CC Region"
                     /label=CC Region
                     /note="CC Region"
     misc_difference complement(2312)
                     /locus_tag="mutated for higher integration efficiency"
                     /replace="g"
                     /label=mutated for higher integration efficiency
                     /note="mutated for higher integration efficiency"
     misc_feature    2534..2553
                     /locus_tag="NBU1-intSeqRev"
                     /label=NBU1-intSeqRev
     misc_feature    2905..2906
                     /locus_tag="deleted?"
                     /label=deleted?
     misc_feature    3008..3038
                     /locus_tag="tet prom putative"
                     /label=tet prom putative
     CDS             3173..5098
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="TetQ"
                     /product="TetQ"
                     /label=TetQ
                     /protein_id="QJT41639.1"
                     /translation="MNIINLGILAHIDAGKTSVTENLLFASGATEKCGCVDNGDTITDS
                     MDIEKRRGITVRASTTSIIWNGVKCNIIDTPGHMDFIAEVERTFKMLDGAVLILSAKEG
                     IQAQTKLLFNTLQKLQIPTIIFINKIDRAGVNLERLYLDIKANLSQDVLFMQNVVDGSV
                     YPVCSQTYIKEEYKEFVCNHDDNILERYLADSEISPADYWNTIIALVAKAKVYPVLHGS
                     AMFNIGINELLDAITSFILPPASVSNRLSSYLYKIEHDPKGHKRSFLKIIDGSLRLRDV
                     VRINDSEKFIKIKNLKTINQGREINVDEVGANDIAIVEDMDDFRIGNYLGAEPCLIQGL
                     SHQHPALKSSVRPDRPEERSKVISALNTLWIEDPSLSFSINSYSDELEISLYGLTQKEI
                     IQTLLEERFSVKVHFDEIKTIYKERPVKKVNKIIQIEVPPNPYWATIGLTLEPLPLGTG
                     LQIESDISYGYLNHSFQNAVFEGIRMSCQSGLHGWEVTDLKVTFTQAEYYSPVSTPADF
                     RQLTPYVFRLALQQSGVDILEPMLYFELQIPQAASSKAITDLQKMMSEIEDISCNNEWC
                     HIKGKVPLNTSKDYASEVSSYTKGLGIFMVKPCGYQITKGGYSDNIRMNEKDKLLFMFQ
                     KSMSSK"
     misc_feature    4009
                     /locus_tag="mut to T for domestication"
                     /label=mut to T for domestication
     CDS             5589..6449
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="beta-lactamase"
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="derived from pBR322"
                     /protein_id="QJT41632.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_difference 6308
                     /locus_tag="misc_difference"
                     /replace="t"
                     /label=misc_difference
     misc_recomb     6488..7173
                     /locus_tag="RP4 oriT/RSK oriV(1)"
                     /label=RP4 oriT/RSK oriV
                     /label=RP4 oriT/RSK oriV(1)
                     /note="RP4 oriT/RSK oriV"
ORIGIN
        1 gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg acacaggaac
       61 acttaacggc tgacatggga attcccctcc accgcggtgt attaatgcgg ctgccccttg
      121 agacggcatc ttccatctac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct
      241 gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcactggcc gtcgttttac
      301 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      361 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      421 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
      481 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca taggatctga
      541 tgcatcgtct caccgataat tgcctatctt ccagtgatgg aacagcattt gtgcattggc
      601 tgcaacaatc agccttactt gtgcctgttc tatttccgaa ccgaccgctt gtatgaatgc
      661 atcaaaattc gttttctcta cgttggattc cttgttgctc atattgtgat gataatttct
      721 acaaatatag tcattggtaa ctatctatga aactgtttga tacttttatc agtctaatag
      781 ttttacaagg tctttcttca tttcttcgtc aatatccctg tatcgtctga aagctttgct
      841 tccctccttg tgtcccgaca gtgcggaaac aaggttcggg tctttcactt ttttatagat
      901 attgccgata aacgtacgtc ttgccagatg gctgcttgcc acttcataaa taggtcgttt
      961 gatttcgttg tgcgtcaacg ggtctaagat tgttacgatg cggtcaactc cagctaattt
     1021 gaatatcttt tttatggcat cattgtactt ttgctcggat atgaacggca acagttttcc
     1081 ctcatattct ttgtagcgtt caaggatttc tttcgctttg tcgttaagtg gaacacgtac
     1141 cgtaaccgga ttcccctctt tggttttctt gggaatatat tctatggctt cattgaccac
     1201 atttagtttg gtcattcggt acaggtcgct caccctgcat cctatcagtg tctgaaatat
     1261 gaatatatcc ctctgtattg ccagttgtgg ggtggcagaa aggtctgcat taaaaatcct
     1321 gtccctttct tcgagtgtta tataataagg tgtaccatat gtacactcct ctatcggaaa
     1381 cttgtcgaaa ggtctgtttg tggtgcgttt gttatcgaag caccacagga agaatgtgcg
     1441 tattcttgaa aaacagtcta tcagcgtgtt tttgcttctg ggctgtggtg tcctcttttc
     1501 gggaatggct tcataaatgc tcgggtaaag ttcataatac tggtattcgt tctgaaagaa
     1561 atcccacata tcccgaagcg tgtcaggtgt taccaaatcc acatcaagga taaagccctt
     1621 ttgtcctctc tttgtagccc ttacatatag ttcataacgc agtaaggctc ttttgacaac
     1681 ccggaaattt ttctttcgta cttccgacaa agggtgcttg tttagaaatt catcgaatag
     1741 ttctccaatg gtaggcttga taaccacttc ctccggcaaa aaatattttt caggatggta
     1801 aaatttatca agtgttgttt ttagccattc tttgtctatt gcttcctttt cttgttgata
     1861 taccttctcg atataggttt tcaactggcg tatctcttca tttatatggg tacgcatttc
     1921 ttcattgcag acagctttcg tttttacaca ttcgtcttta tcatcccata gattgggatt
     1981 gatggctaac tgagtaggag caacggagtc taactgcctt ccgtttctaa aacggacata
     2041 gattgtagcc atggattctg tatcatatcg tttggctgct tttttaatga taaaggttac
     2101 tttcatagac tttcaggttg aattttactc tgctgcaaat ataaatattt tccccagcat
     2161 tttccccaca tctgctaaat attttgcaat tcgattaaac ttggattaaa atttaatagg
     2221 attataacat attgaaatac agtgtaattg tggcgttttt ccgcattttt ctttttaccc
     2281 acattttccc cactttaatt atttcaaatg cggggtctgg gtacaagaaa gaaagctaag
     2341 tatttgatag ttcaatactt agctttttct tttgcttgaa ttttccccac attttcccca
     2401 cacgtgcaaa aaatatagca gtaagtcatt atttcttttg gttgaacgta gagagtagcg
     2461 atattaaaag aatccgatga gaaaagacta atatttatct atccattcag tttgattttt
     2521 caggacttta catcgtcctg aaagtatttg ttggtaccgg taccgaggac gcgtaaacat
     2581 ttacagttgc atgtggccta ttgtttagga cgcgttatct ccttaacgta cgttttcgtt
     2641 ccattggccc tcaaaccccg ttatatacat tcatgtccat ttatgtaaaa aatcctgctg
     2701 accttgttta tgtcttgtca gtcaccattt gcaaaaccat atttgaccct caaagaggct
     2761 gaatttgata agcaacttgc tacatactca taataaggag ctaaatagaa cacgaatggg
     2821 aaatactcaa atgccaaact aaagaagata ttggccaaaa taaacgctat accgagagag
     2881 aaacttgatt tttcaacttc ctaaccaaca gtgttgttca aacatttcta cttatttgta
     2941 cttaccagtt gaacctacgt ttccctaata aaatgtctat ggtaaaaagt taaaaaatcc
     3001 tcctactttt gttagatata tttttttgtg taattttgta atcgttatgc ggcagtaata
     3061 atatacatat taatacgagt taggaatcct gtagttctca tatgctacga ggaggtatta
     3121 aaaggtgcgt ttcgacaatg catctattgt agtatattat tgcttaatcc aaatgaatat
     3181 tataaattta ggaattcttg ctcacattga tgcaggaaaa acttccgtaa ccgagaatct
     3241 gctgtttgcc agtggagcaa cggaaaagtg cggctgtgtg gataatggtg acaccataac
     3301 ggactctatg gatatagaga aacgtagagg aattactgtt cgggcttcta cgacatctat
     3361 tatctggaat ggtgtgaaat gcaatatcat tgacactccg ggacacatgg attttattgc
     3421 ggaagtggag cggacattca aaatgcttga tggagcagtc ctcatcttat ccgcaaagga
     3481 aggcatacaa gcgcagacaa agttgctgtt caatacttta cagaagctgc aaatcccgac
     3541 aattatattt atcaataaga ttgaccgagc cggtgtgaat ttggagcgtt tgtatctgga
     3601 tataaaagca aatctgtctc aagatgtcct gtttatgcaa aatgttgtcg atggatcggt
     3661 ttatccggtt tgctcccaaa catatataaa ggaagaatac aaagaatttg tatgcaacca
     3721 tgacgacaat atattagaac gatatttggc ggatagcgaa atttcaccgg ctgattattg
     3781 gaatacgata atcgctcttg tggcaaaagc caaagtctat ccggtgctac atggatcagc
     3841 aatgttcaat atcggtatca atgagttgtt ggacgccatc acttctttta tacttcctcc
     3901 ggcatcggtc tcaaacagac tttcatctta tctttataag atagagcatg accccaaagg
     3961 acataaaaga agttttctaa aaataattga cggaagtctg agacttcgag acgttgtaag
     4021 aatcaacgat tcggaaaaat tcatcaagat taaaaatcta aaaactatca atcagggcag
     4081 agagataaat gttgatgaag tgggcgccaa tgatatcgcg attgtagagg atatggatga
     4141 ttttcgaatc ggaaattatt taggtgctga accttgtttg attcaaggat tatcgcatca
     4201 gcatcccgct ctcaaatcct ccgtccggcc agacaggccc gaagagagaa gcaaggtgat
     4261 atccgctctg aatacattgt ggattgaaga cccgtctttg tccttttcca taaactcata
     4321 tagtgatgaa ttggaaatct cgttatatgg tttaacccaa aaggaaatca tacagacatt
     4381 gctggaagaa cgattttccg taaaggtcca ttttgatgag atcaagacta tatacaaaga
     4441 acgacctgta aaaaaggtca ataagattat tcagatcgaa gtgccgccca acccttattg
     4501 ggccacaata gggctgactc ttgaaccctt accgttaggg acagggttgc aaatcgaaag
     4561 tgacatctcc tatggttatc tgaaccattc ttttcaaaat gccgtttttg aagggattcg
     4621 tatgtcttgc caatccgggt tacatggatg ggaagtgact gatctgaaag taacttttac
     4681 tcaagccgag tattatagcc cggtaagtac acctgctgat ttcagacagc tgacccctta
     4741 tgtcttcagg ctggccttgc aacagtcagg tgtggacatt ctcgaaccga tgctctattt
     4801 tgagttgcag ataccccaag cggcaagttc caaagctatt acagatttgc aaaaaatgat
     4861 gtctgagatt gaagacatca gttgcaataa tgagtggtgt catattaaag ggaaagttcc
     4921 attaaataca agtaaagact atgcatcaga agtaagttca tacactaagg gcttaggcat
     4981 ttttatggtt aagccatgcg ggtatcaaat aacaaaaggc ggttattctg ataatatccg
     5041 catgaacgaa aaagataaac ttttattcat gttccaaaaa tcaatgtcat caaaataatg
     5101 gtataacctc tccttaattt attgcatctc ttttcgaata tttatgtttt ttgagaaaag
     5161 aacgtactca tggttcatcc cgatatgcgt atcggtctgt atatcagcaa ctttctatgt
     5221 gtttcaacta caatagtcat ctattctcat ctttctgagt ccaccccctg caaagcccct
     5281 ctttacgaca taaaaattcg gtcggaaaag gtatgcaaaa gatgtttctc tctttaagag
     5341 aaactcttcg ggatgcaaaa atatgaaaat aactccaatt caccaaatta tatagcgact
     5401 tttttacaaa atgctaaaat ttgttgattt ccgtcaagca attgttgagc aaaaatgtct
     5461 tttacgataa aatgatacct caatatcaac tgtttagcaa aacgatattt ctcttaaaga
     5521 gagaaacacc tttttgttca ccaatccccg acttttaatc ccgcggccat gattgaaaaa
     5581 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5701 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5761 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5881 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6001 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6121 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6181 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6301 gtgggtcacg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6421 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca taacgcgtca
     6481 attcgagggg gatcaattcc gtgataggtg ggctgccctt cctggttggc ttggtttcat
     6541 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     6601 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     6661 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     6721 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     6781 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaacggaatt gatccggcca
     6841 cgatgcgtcc ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact
     6901 aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa
     6961 cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat
     7021 gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa
     7081 tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt
     7141 ttaaggttta acggttgtgg acaacaagcc agg
//
