LOCUS       Exported                6719 bp DNA     circular SYN 20-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6719)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6719)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6719)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4578..4580
                     /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(142..819)
                     /codon_start=1
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the
                     lambda
                     Red system"
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(819..1601)
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             complement(2615..3472)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3473..3544)
                     /label=AmpR promoter
     CDS             complement(3660..4370)
                     /codon_start=1
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda 
                     repressor"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     misc_feature    complement(4482..4500)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(5023..5970)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(6018..6240)
                     /direction=LEFT
                     /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 cggaagtaaa
      541 ttcaaacagg gttctggcgt cgttctcgta ctgttttccc caggccagtg ctttagcgtt
      601 aacttccgga gccacaccgg tgcaaacctc agcaagcagg gtgtggaagt aggacatttt
      661 catgtcaggc cacttctttc cggagcgggg ttttgctatc acgttgtgaa cttctgaagc
      721 ggtgatgacg ccgagccgta atttgtgcca cgcatcttcc 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 cattgctaca 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 ggcgtcaata 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 tctaagctca
     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 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat
     4621 gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata
     4681 agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc
     4741 aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt
     4801 gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca
     4861 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     4921 atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt
     4981 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     5041 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     5101 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     5161 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     5221 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     5281 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     5341 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     5401 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     5461 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     5521 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     5581 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     5641 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     5701 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     5761 taatacacca taagcatttt ccctactgat gttcatcatc tgaacgtatt ggttataagt
     5821 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     5881 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     5941 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     6001 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     6061 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     6121 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     6181 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     6241 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     6301 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     6361 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     6421 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     6481 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     6541 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     6601 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     6661 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtc
//
