LOCUS       40924_40372        4781 bp DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4781)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4781)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(536..739)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(740..1043)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(1165..1183)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1195..1213)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1233..1249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1257..1273)
                     /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(1281..1311)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1326..1347)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1635..2223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2397..3254)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3255..3359)
                     /label=AmpR promoter
     polyA_signal    complement(3447..3528)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3761..4357)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(4454..4770)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctagcggcc gcatacaaaa
      421 aaccaacaca cagatccaat gaaaataaaa gatcctttat taagcttact accgttgtta
      481 taggtgtctc ttgaacacct ataacaacgg tagtggatcc acggttcact aaaccagctc
      541 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
      601 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
      661 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
      721 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
      781 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
      841 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
      901 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
      961 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     1021 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     1081 taattgatta ctattaataa ctaagatctg gtaccttgaa ttcatgcttc tcctcccttt
     1141 agtgagggta attctctctc tctccctata gtgagtcgta ttaattcctt ctcttctata
     1201 gtgtcaccta aatcgttgca attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     1261 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1321 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1381 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1441 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1501 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1561 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1801 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2041 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa aaaaaccacc
     2161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2221 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2281 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2341 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2401 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2461 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2521 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2581 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2641 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2701 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2761 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2821 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2881 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2941 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3001 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3061 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3121 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3181 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3241 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3301 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3361 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3421 tataaaaata ggcgtatcac gagattgcag tgaaaaaaat gctttatttg tgaaatttgt
     3481 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3541 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3601 aacctctaca aatgtggtat ggctgattat gatcctctag agtcggtggg cctcgggggc
     3661 gggtgcgggg tcggcggggc cgccccgggt ggcttcggtc ggagccatgg ggtcgtgcgc
     3721 tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca
     3781 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     3841 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
     3901 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     3961 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     4021 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     4081 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     4141 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     4201 agagttcttg cagctcggtg acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg
     4261 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc
     4321 gggtggcgag gcgcaccgtg ggcttgtact cggtcatgga aggtcgtctc cttgtgaggg
     4381 gtcaggggcg tgggtcaggg gatggtggcg gcaccggtcg tggcggccga cctgcaggca
     4441 tgcaagctag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     4501 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     4561 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     4621 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     4681 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     4741 ctgctgggga gcctggggac tttccacacc aatctttcgt c
//
