LOCUS       40924_41495        4354 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4354)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4354)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(759..1355)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(1380..1879)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     protein_bind    2090..2108
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2120..2136)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2166..2184)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2205..2221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2229..2245)
                     /label=lac operator
                     /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(2253..2283)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2298..2319)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2607..3195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3369..4226)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4227..4331)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcatgggg tcgtgcgctc
      721 ctttcggtcg ggcgctgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg
      781 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
      841 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
      901 tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
      961 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     1021 aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     1081 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     1141 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     1201 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     1261 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     1321 gtggcgaggc gcaccgtggg cttgtactcg gtcatggtaa gctagcttgg gctgcaggtc
     1381 gaaaggcccg gagatgagga agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg
     1441 cagaatgccg ggcctccgga ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg
     1501 agcccgcccc cggacccacc ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc
     1561 tgctattggc cgctgcccca aaggcctacc cgcttccatt gctcagcggt gctgtccatc
     1621 tgcacgagac tagtgagacg tgctacttcc atttgtcacg tcctgcacga cgcgagctgc
     1681 ggggcggggg ggaacttcct gactagggga ggagtagaag gtggcgcgaa ggggccacca
     1741 aagaacggag ccggttggcg cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca
     1801 cttgtgtagc gccaagtgcc cagcggggct gctaaagcgc atgctccaga ctgccttggg
     1861 aaaagcgcct cccctacccg gtagaattcg aacgctgacg tcatcaaccc gctccaagga
     1921 atcgcgggcc cagtgtcact aggcgggaac acccagcgcg cgtgcgccct ggcaggaaga
     1981 tggctgtgag ggacagggga gtggcgccct gcaatatttg catgtcgcta tgtgttctgg
     2041 gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt ccctatcagt
     2101 gatagagatc taagcttatc gataccgtcg acctcgaggg ggggcccggt acccagcttt
     2161 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2221 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2281 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2341 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2401 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2461 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2521 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2581 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2641 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2701 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2761 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2821 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2881 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2941 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3001 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3061 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3121 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3181 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3241 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3301 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3361 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3421 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3481 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3541 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3601 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3661 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3721 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3781 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3841 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3901 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3961 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4021 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4081 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4141 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4201 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4261 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4321 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
