LOCUS       Exported                6294 bp ds-DNA     circular SYN 20-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  AUTHORS   Kaiyi Jiang
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 13, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6294
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        80..98
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     157..191
                     /label=Primer 1
     CDS             159..1595
                     /codon_start=1
                     /label=Feature 14
                     /translation="MKRKREQMTLWKAAFVNGQETFKSWIDKARMLELNCDVSSASSTH
                     YSDLNLKTKCAKMEDKFMCTFSVGIRPTSKQKRTLNQMLKVSNHAYNWCNYLVKEKDFK
                     PKQFDLQRVVTKTNSTDVPAEYRLPGDDWFFDNKMSSIKLTACKNFCTMYKSAQTNQKK
                     TKVDLRNKDIAMLREGSFEVQKKYVRLLTEKDIPDERIRQSRIALMADNFSKSKKDWKE
                     RFLRLSKNVSKIPPLSHDMKVCKRPNGKFVLQIPCDPIYTRQIQVHTSDSICSIDPGGR
                     TFATCYDPSNIKAFQIGPEADKKEVIHKYHEKIDYVHRLLAYAQKKKQTQAVQDRIGQL
                     KKLHLKLKTYVDDVHLKLCSYLVKNYKLVVLGKISVSSIVRKDRPNHLAKKANRDLLCW
                     QHYRFRQRLLHRVRGTDCEAIAQDERYTSKTCGNCGVKNNKLGGKETFICESCNYKTHR
                     DVNGARNILCKYLGLFPFAA"
     misc_feature    336..371
                     /label=WED
     misc_feature    372..695
                     /label=rec
     misc_feature    696..938
                     /label=wed
     misc_feature    969..1589
                     /label=ruvc
     primer_bind     complement(1569..1595)
                     /label=Primer 2
     terminator      1673..1720
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1819..1828)
                     /label=PD9440
     promoter        complement(2086..2114)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2135..2151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2159..2175
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2183..2213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2228..2249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2907..2925
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(3473..3491)
                     /label=Cre-Rev
     rep_origin      complement(3935..4523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4694..5554)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5555..5659)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6135..6151
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
       61 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      121 agaaataatt ttgtttaact ttaagaagga gatataccat gaagcggaaa agagaacaaa
      181 tgactttgtg gaaggcggcg tttgtaaacg gccaagagac atttaaatca tggattgata
      241 aggctcgtat gcttgaactt aattgcgacg ttagctccgc atccagtact cattattctg
      301 atcttaacct gaagacaaaa tgtgctaaaa tggaggacaa gttcatgtgt acattcagtg
      361 ttggcatcag accaacatcc aagcaaaaga gaacccttaa ccagatgtta aaggtatcta
      421 accacgccta taactggtgt aattatttag ttaaagagaa agatttcaaa ccaaagcagt
      481 tcgaccttca gcgcgttgta actaaaacga atagtactga tgtaccagct gaatatagat
      541 tgccaggaga tgactggttt tttgacaata agatgtcatc aataaagttg accgcatgca
      601 agaacttctg taccatgtac aaaagtgctc agaccaacca aaagaagacc aaagtcgacc
      661 ttagaaataa agatatcgca atgttacgtg agggatcgtt tgaagtgcaa aagaagtatg
      721 ttagactttt gacagagaag gacatacccg acgagcggat ccgtcagagt agaatagcgc
      781 tgatggcaga taatttttct aaatctaaaa aagactggaa agaacggttt ctgcggttaa
      841 gtaaaaatgt ctcaaagata cctccgctgt cccacgacat gaaagtatgc aaacggccga
      901 acggaaagtt cgtcctgcag atcccttgcg accctattta taccagacaa atccaggtgc
      961 atacgtctga tagtatctgc agcattgacc cagggggacg gacgtttgct acgtgctatg
     1021 acccaagcaa cattaaagcg tttcagattg ggcctgaagc cgacaagaaa gaagttattc
     1081 ataaatatca cgagaagata gactatgttc accgtctttt ggcgtacgct caaaagaaaa
     1141 agcaaacgca agcggtgcaa gatcgtattg gacagcttaa gaaacttcat ttaaagttaa
     1201 aaacctacgt tgatgacgtc catcttaagt tgtgcagtta ccttgtcaaa aattataagc
     1261 tggtcgtcct gggcaagatc agcgtcagca gtatcgttcg caaggacaga cccaatcatc
     1321 ttgccaagaa agccaataga gacttgttat gttggcagca ttatcggttc cggcaaagac
     1381 tgcttcaccg ggttcgcgga accgactgtg aggcaatagc tcaagacgaa cgctacacat
     1441 ccaagacctg tggaaactgt ggggttaaaa acaataaact tggtgggaag gagaccttta
     1501 tctgcgaatc ctgcaattac aaaacccacc gggatgtaaa tggtgcccgc aatattttat
     1561 gtaagtatct tggccttttc cctttcgctg cttaactcga gccttaggag atccggctgc
     1621 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1681 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1741 cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     1801 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     1861 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     1921 tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     1981 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     2041 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     2101 tgataagctg tcaaacatga gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     2161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2761 tgtasngtck cttatacaca tctccgmtsy swssasrmac mymscgwtca gcccgaccgc
     2821 tgcgccttat ccgggaacta gcgtcttgag tccaacccgg taagacacga cttakcgcca
     2881 ctggcagcag ccactggtaa caggatctgt ctcttataca catctccgag cccacgagac
     2941 acatagcgat ctcgtatgcc gtcttctgct tgaaaaaaaa aaamayataa asatcgtcwg
     3001 tatgttagtg ttatgtattc aattgtctgt gtttttgtgt atattagttg tgtttatagt
     3061 tgtgacagaa aaaaaaagac gaacaaaaca aacgaaaaaa aattgaataa accaagggaa
     3121 agaaaaaaaa ctaacaaaaa agatcattag ggcagcgcaa gatgcgtagc agagacagat
     3181 aaatcccgaa caaagccaca aatccgccaa aaccatctga tgattcctac gacatgccta
     3241 gtgtctatac atgcgtgttc cgcggacctc acacaccgaa aaaaatactt gggtatgtgg
     3301 ttgtgcgtga tcaaacgatg atataaaacg aggagagatg aaatagagag ataggaccta
     3361 gagcttttca gtcgggacac ctgtcttgcc agctgcctta atgaatcggc caacgcgcgg
     3421 ggcgaggcgg atttcggaat tggcgctcct cccctctttt atacccacat cgccctgacc
     3481 aggagctccc ttgtgtcgat ctcgggtggt ccccaccagt ctcctgtcac gcttgtctca
     3541 aggtcataat gttcaacaag aaaacctggg tgctatacat gttgcatgtc gttttttttt
     3601 tttcaagcag aagacggcat acgagatcgc tatgtgtctc gtgggctcgg agatgtgtat
     3661 aagagacmgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3721 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3781 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4561 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4621 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4681 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4741 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4801 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4861 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4921 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4981 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5041 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5101 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5161 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5221 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5281 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5341 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5401 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5461 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5521 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5581 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5641 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5701 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5761 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5821 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5881 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5941 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6001 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6061 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6121 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcaagg agatggcgcc
     6181 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     6241 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgcc
//
