LOCUS       40924_10271        6662 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 6662)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6662)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1028..1600
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        1655..1673
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1684..2304
                     /label=TetR
                     /note="tetracycline repressor TetR"
     polyA_signal    2346..2467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2530..2571
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             2581..2598
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    2627..2851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2897..3325
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3339..3668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3716..3763
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3782..4177
                     /label=BSD
                     /note="blasticidin S deaminase"
     polyA_signal    4338..4459
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4508..4524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4532..4548)
                     /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(4556..4586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4601..4622)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4910..5495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5669..6526)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6527..6631)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta cccggggatc ctctagggcc tctgagctat tccagaagta
      961 gtgaagaggc ttttttggag gcctaggctt ttgcaaaaag ctccggatcg atcctgagaa
     1021 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     1081 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     1141 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     1201 gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt
     1261 gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc
     1321 actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac
     1381 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     1441 aaatattctt attggtagaa acaactacat cctggtcatc atcctgcctt tctctttatg
     1501 gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc
     1561 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1621 tattgtgctg tctcatcatt ttggcaaaga attgtaatac gactcactat agggcgaatt
     1681 gatatgtcta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     1741 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1801 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     1861 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1921 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     1981 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     2041 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
     2101 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     2161 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     2221 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     2281 caacttaaat gtgaaagtgg gtccgcgtac agcggatccc gggaattcag atcttattaa
     2341 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2401 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2461 tatcttatca tgtctggtcg aatccatcac actggcggcc gctcgagtct agagggcccg
     2521 cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt
     2581 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     2641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2821 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2881 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3301 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3421 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3601 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3661 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3721 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3781 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     3841 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     3901 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     3961 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     4021 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     4081 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     4141 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg gccgaggagc
     4201 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4261 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4321 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4381 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4441 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4501 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4561 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4621 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4681 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4741 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4801 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4861 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4921 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4981 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5041 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5101 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5161 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5221 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5281 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5341 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5401 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6661 tc
//
