LOCUS       Exported                4703 bp DNA     circular SYN 16-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4703)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             954..977
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     polyA_signal    1088..1222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1358..1568
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     primer_bind     complement(1596..1612)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1825..2280
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2562..2666
                     /label=AmpR promoter
     CDS             2667..3524
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3698..4286
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4574..4595
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4610..4640
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4648..4664
                     /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)."
     primer_bind     4672..4688
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attctcttca ccaaggccat tcgcgccacc atggattaca
      961 aggacgacga tgacaagaga tctcaatggt ggtggtggtg ggtcgacgta cccgggcgtc
     1021 aagcttgcta gatatccgca tgcggccgct aagtaagtaa ggatccccag cttggccgcc
     1081 atggcccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1141 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1201 tgtatcttat catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac
     1261 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     1321 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1381 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     1441 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     1501 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     1561 cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt
     1621 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1681 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1741 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     1801 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1861 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1921 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1981 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2041 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2101 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2161 tctcgggcta ttcttttaat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2221 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2281 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2341 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2401 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2461 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2521 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2581 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2641 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2701 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2761 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2821 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2881 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2941 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3001 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3061 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3121 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3181 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3241 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3301 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3361 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3421 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3481 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3541 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3601 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3661 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3721 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3781 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3841 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3901 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3961 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4021 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4081 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4141 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4201 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4261 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4321 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4381 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4441 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     4501 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     4561 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     4621 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     4681 gctatgacat gattacgaat taa
//
