LOCUS       Exported                4645 bp DNA     circular SYN 08-APR-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4645)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4645)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     508..524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..1292
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             1449..1475
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     CDS             1500..2216
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2224..2240
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHH"
     primer_bind     complement(2554..2569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2577..2593)
                     /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(2601..2631)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2646..2667)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3717..4574)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(4575..4645,1..34))
                     /label=AmpR promoter
ORIGIN
        1 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
       61 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      121 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      181 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      241 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      301 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      361 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      421 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      481 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgcc
      541 tgcagtcaca tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc
      601 tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc
      661 ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc
      721 tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac
      781 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
      841 accacgtctt caaagcaagt ggattgatgt gataacatgg tggagcacga cacacttgtc
      901 tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga gacttttcaa
      961 caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt
     1021 gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
     1081 gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
     1141 agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat
     1201 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     1261 atataaggaa gttcatttca tttggagagg acctcgacct caacacaaca tatacaaaac
     1321 aaacgaatct caagcaatca agcattctac ttctattgca gcaatttaaa tcatttcttt
     1381 taaagcaaaa gcaattttct gaaaattttc accatttacg aacgatactc gagatgaccc
     1441 gtggttctca tcaccatcac catcaccatc accatcacgc catggtcgac gtactagtga
     1501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1681 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1741 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1921 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1981 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2041 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2101 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2161 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcta
     2221 gacatcacca tcaccatcac ggatcctgag cggccgccac cgcggtggag ctcgaatttc
     2281 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     2341 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     2401 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     2461 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     2521 tctatgttac tagatcggga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     2581 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2641 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2761 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4621 ctcatgagcg gatacatatt tgaat
//
