LOCUS       無題_2        6314 bp DNA     circular SYN 09-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 9, 2023 from SnapGene Viewer 6.1.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..673
                     /label=chicken beta-actin promoter
     intron          674..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1744..1773
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1807..2904
                     /label=mouse TSSK1
     polyA_signal    3061..3116
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     promoter        complement(3470..3488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3498..3514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3656..4111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4137..4241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4242..5102
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5273..5861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6149..6170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6185..6215
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6223..6239
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     6247..6263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6284..6302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcatcg atctcgacat tgattattga ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatgggtcg aggtgagccc cacgttctgc
      421 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
      481 ttattttgtg cagcgatggg ggcggggggg gggggggcgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
      841 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
      901 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     1021 gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg ctgtaacccc
     1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1201 gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1261 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca gcctcggggc
     1561 tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaatc tagcgccgcc
     1741 atggaacaaa aactcatctc agaagaggat ctggggggcg ggtctagagg atccctgcag
     1801 gaattcatgg atgacgctgc cgtcctcaag cgacgaggct acatcatggg gataaacttg
     1861 ggagagggct catatgccaa agtcaaatcc gcttactctg agcgcctaaa gttcaacgtg
     1921 gcggtcaaga tcatcgatcg caagaaagcc ccctcagact tcctggagaa attccttccc
     1981 cgggagattg agattctggc catgctaaac caccgctcta ttgtcaagac ctatgagatc
     2041 tttgagacat cagatggcaa ggtctacatt gtcatggagc ttggggtcca gggtgacctc
     2101 ctcgagttca tcaaaaccag aggagccctg caagaggatg atgctcgaaa gaagttccac
     2161 cagctctcct cggccatcaa gtactgccac gacctggatg tcgtccaccg agacctcaaa
     2221 tgtgagaacc ttctgcttga caaggacttc aacatcaaac tgtctgactt cggcttctcc
     2281 aagcgctgct tgcgggatga tagtggtcga ctgatattaa gcaagacctt ctgtgggtca
     2341 gcagcttatg cggcccccga ggtgctgcag ggcatcccct accagcccaa ggtgtatgac
     2401 atctggagcc tgggtgtgat tctctacatc atggtctgtg gctccatgcc ctacgatgac
     2461 tccaacatca aaaagatgct gcgcatccag aaggagcacc gtgtcaactt tccacgctct
     2521 aaacacctga ctggtgagtg caaggacctc atctacagaa tgttacagcc agatgtcaac
     2581 cgccggctgc acatcgatga gatccttaac cactgctggg tgcagcccaa ggcacgaggt
     2641 ctgtcctctg gagccatcaa caaggagggg gaaagctccc gggccactga gccctcatgg
     2701 atccctgaac ctggtgctga taagaagtct gccaccaagt tggagcctag ggaagaggca
     2761 cgatctgagg cccggtctga gtcaaaaccc caggaagata ctttgcaggt ggtcaggcag
     2821 tcagagaacg tgggcctcag cagcgaactg aacagggaca cagaggaagg gcacccccaa
     2881 cagccttcag agacacatac ttaggcggcc gcctcgagac tcctcaggtg caggctgcct
     2941 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     3001 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     3061 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     3121 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     3181 tggcaacata tgccatatgc tggctgccat gaacaaaggt ggctataaag aggtcatcag
     3241 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     3301 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     3361 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     3421 cctcttctct tatgaagatc cctcgactta attaaggtac ccaattcgcc ctatagtgag
     3481 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3541 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3601 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4081 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4921 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5221 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5401 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5641 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6121 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6181 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6241 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     6301 gggaacaaaa gctg
//
