pSIM6

LOCUS       pSIM6                6716 bp DNA     circular SYN 12-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  AUTHORS   Biofeng
  TITLE     Direct Submission
  JOURNAL   Exported Jul 12, 2023 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    4..22
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(139..819)
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(816..1601)
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     protein_bind    2034..2048
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     CDS             complement(2612..3472)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3473..3544)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3657..4370)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             4459..4467
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    4482..4500
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(5017..5967)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6015..6237
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cagctgtctc ttatacacat ctccgctgtg ctttcagtgg atttcggata acagaaaggc
       61 cgggaaatac ccagcctcgc tttgtaacgg agtagacgaa agtgattgcg cctacccgga
      121 tattatcgtg aggatgcgtc atcgccattg ctccccaaat acaaaaccaa tttcagccag
      181 tgcctcgtcc attttttcga tgaactccgg cacgatctcg tcaaaactcg ccatgtactt
      241 ttcatcccgc tcaatcacga cataatgcag gccttcacgc ttcatacgcg ggtcatagtt
      301 ggcaaagtac caggcatttt ttcgcgtcac ccacatgctg tactgcacct gggccatgta
      361 agctgacttt atggcctcga aaccaccgag ccggaacttc atgaaatccc gggaggtaaa
      421 cgggcatttc agttcaaggc cgttgccgtc actgcataaa ccatcgggag agcaggcggt
      481 acgcatactt tcgtcgcgat agatgatcgg ggattcagta acattcacgc cggaagtgaa
      541 ctcaaacagg gttctggcgt cgttctcgta ctgttttccc caggccagtg ctttagcgtt
      601 aacttccgga gccacaccgg tgcaaacctc agcaagcagg gtgtggaagt aggacatttt
      661 catgtcaggc cacttctttc cggagcgggg ttttgctatc acgttgtgaa cttctgaagc
      721 ggtgatgacg ccgagccgta atttgtgcca cgcatcatcc ccctgttcga cagctctcac
      781 atcgatcccg gtacgctgca ggataatgtc cggtgtcatg ctgccacctt ctgctctgcg
      841 gctttctgtt tcaggaatcc aagagctttt actgcttcgg cctgtgtcag ttctgacgat
      901 gcacgaatgt cgcggcgaaa tatctgggaa cagagcggca ataagtcgtc atcccatgtt
      961 ttatccaggg cgatcagcag agtgttaatc tcctgcatgg tttcatcgtt aaccggagtg
     1021 atgtcgcgtt ccggctgacg ttctgcagtg tatgcagtat tttcgacaat gcgctcggct
     1081 tcatccttgt catagatacc agcaaatccg aaggccagac gggcacactg aatcatggct
     1141 ttatgacgta acatccgttt gggatgcgac tgccacggcc ccgtgatttc tctgccttcg
     1201 cgagttttga atggttcgcg gcggcattca tccatccatt cggtaacgca gatcggatga
     1261 ttacggtcct tgcggtaaat ccggcatgta caggattcat tgtcctgctc aaagtccatg
     1321 ccatcaaact gctggttttc attgatgatg cgggaccagc catcaacgcc caccaccgga
     1381 acgatgccat tctgcttatc aggaaaggcg taaatttctt tcgtccacgg attaaggccg
     1441 tactggttgg caacgatcag taatgcgatg aactgcgcat cgctggcatc acctttaaat
     1501 gccgtctggc gaagagtggt gatcagttcc tgtgggtcga cagaatccat gccgacacgt
     1561 tcagccagct tcccagccag cgttgcgagt gcagtactca ttcgttttat acctctgaat
     1621 caatatcaac ctggtggtga gcaatggttt caaccatgta ccggatgtgt tctgccatgc
     1681 gctcctgaaa ctcaacatcg tcatcaaacg cacgggtaat ggattttttg ctggccccgt
     1741 ggcgttgcaa atgatcgatg catagcgatt caaacaggtg ctggggcagg cctttttcca
     1801 tgtcgtctgc cagttctgcc tctttctctt cacgggcgag ctgctggtag tgacgcgccc
     1861 agctctgagc ctcaagacga tcctgaatgt aataagcgtt catggctgaa ctcctgaaat
     1921 agctgtgaaa atatcgcccg cgaaatgccg ggctgattag taatccggaa tcgcacttac
     1981 ggccaatgct tcgtttcgta tcacacaccc caaagccttc tgctttgaat gctgcccttc
     2041 ttcagggctt aatttttaag agcgtcacct tcatggtggt cagtgcgtcc tgctgatgtg
     2101 ctcagtatca ccgccagtgg tatttatgtc aacaccgcca gagataattt atcaccgcag
     2161 atggttatct gtatgttttt tatatgaatt tattttttgc aggggggcat tgtttggtag
     2221 gtgagagatc tgaattgcta tgtttagtga gttgtatcta tttatttttc aataaataca
     2281 attggttatg tgttttgggg gcgatcgtga ggcaaagaaa acccggcgct gaggccgggt
     2341 taagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     2941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     3241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3541 aatgaattca atccatttac tatgttatgt tctgagggga gtgaaaattc ccctaattcg
     3601 atgaagattc ttgctcaatt gttatcagct atgcgccgac cagaacacct tgccgatcag
     3661 ccaaacgtct cttcaggcca ctgactagcg ataactttcc ccacaacgga acaactctca
     3721 ttgcatggga tcattgggta ctgtgggttt agtggttgta aaaacacctg accgctatcc
     3781 ctgatcagtt tcttgaaggt aaactcatca cccccaagtc tggctatgca gaaatcacct
     3841 ggctcaacag cctgctcagg gtcaacgaga attaacattc cgtcaggaaa gcttggcttg
     3901 gagcctgttg gtgcggtcat ggaattacct tcaacctcaa gccagaatgc agaatcactg
     3961 gcttttttgg ttgtgcttac ccatctctcc gcatcacctt tggtaaaggt tctaagctta
     4021 ggtgagaaca tccctgcctg aacatgagaa aaaacagggt actcatactc acttctaagt
     4081 gacggctgca tactaaccgc ttcatacatc tcgtagattt ctctggcgat tgaagggcta
     4141 aattcttcaa cgctaacttt gagaattttt gtaagcaatg cggcgttata agcatttaat
     4201 gcattgatgc cattaaataa agcaccaacg cctgactgcc ccatccccat cttgtctgcg
     4261 acagattcct gggataagcc aagttcattt ttcttttttt cataaattgc tttaaggcga
     4321 cgtgcgtcct caagctgctc ttgtgttaat ggtttctttt ttgtgctcat acgttaaatc
     4381 tatcaccgca agggataaat atctaacacc gtgcgtgttg actattttac ctctggcggt
     4441 gataatggtt gcatgtacta aggaggttgt atggaacaac gagatgtgta taagagacag
     4501 ctgacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat
     4561 cgcagatccg gcttcaggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
     4621 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
     4681 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     4741 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     4801 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     4861 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     4921 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     4981 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     5041 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     5101 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     5161 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     5221 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     5281 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     5341 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     5401 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     5461 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     5521 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     5581 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     5641 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     5701 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     5761 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     5821 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     5881 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     5941 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     6001 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     6061 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     6121 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     6181 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     6241 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     6301 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     6361 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     6421 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     6481 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     6541 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     6601 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
     6661 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtc
//