LOCUS       Exported                6565 bp ds-DNA     circular SYN 13-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   Richard Davis
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 17, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6565
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     promoter        253..463
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        559..1736
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_recomb     1754..1795
                     /label=attB
                     /note="from myobacteriumphage bxb1 (Thomson et al 2012)"
     protein_bind    1802..1835
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        1877..2675
                     /label=CBh
                     /note="CBh"
     regulatory      2682..2690
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2688..3386
                     /codon_start=1
                     /label=mStayGold
                     /translation="MRGSDPMVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSH
                     EGSHKGKYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQ
                     YKGDGSIHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGV
                     ECTVTLQYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHI
                     IQSETLEAHL"
     polyA_signal    3398..3622
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3654..3686
                     /label=lox251
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     rep_origin      3923..4378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4660..4764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4765..5625
                     /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      5796..6384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctta
      241 attccctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttgc
      481 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      541 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      601 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      661 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      721 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      781 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      841 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      901 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      961 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1021 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1081 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1141 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1201 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1261 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1321 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1381 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1441 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1501 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1561 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1621 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1681 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1741 ttcgccacca tggccggctt gtcgacgacg gcggtctccg tcgtcaggat catccggatc
     1801 cataacttcg tatagcatac attatacgaa gttatcatga tattcggcaa gcaggcatcg
     1861 actagttaat taagcccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     1921 acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat
     1981 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2041 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca
     2101 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2161 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     2221 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
     2281 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
     2341 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     2401 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc
     2461 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     2521 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg
     2581 agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg
     2641 gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg cggggttctg
     2701 atccgatggt gtctacaggc gaggagctgt ttaccggcgt ggtgcccttc aagttccagc
     2761 tgaagggcac catcaacggc aagagcttca ccgtggaagg cgagggcgag ggcaatagcc
     2821 acgagggcag ccacaaaggc aagtacgtgt gcaccagcgg caaactgcca atgtcttggg
     2881 ccgccctggg aactagcttc ggctatggca tgaagtacta caccaagtac cccagcggcc
     2941 tgaagaactg gttccacgag gtgatgcccg agggcttcac ctacgacaga cacatccagt
     3001 acaagggcga cggcagcatc cacgccaagc accagcactt catgaagaac ggcacctacc
     3061 acaacatcgt ggagttcacc ggccaggact tcaaggagaa cagccccgtg ctgaccggcg
     3121 acatggacgt gagcctgccc aacgaggtgc agcacatccc catagatgac ggcgtggagt
     3181 gcacagtgac cctgcagtac cctctgctga gcgacgagag caagtgcgtg gaggcctacc
     3241 agaacaccat catcaagccc ctgcacaatc agccagcccc cgatgtgcca ttccactgga
     3301 tcagaaagca gtacacccag agcaaggacg acaccgagga gagggaccac atcatccaga
     3361 gcgagacact ggaggcccac ctgtaacaac gttgcacctg tgccttctag ttgccagcca
     3421 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3481 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3541 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3601 ggggatgcgg tgggctctat ggcgctcgct cgctgcaaga actcttcctc acgataactt
     3661 cgtataggag actttatacg aagttaagcg ctcactggcc gtcgttttac aacgtcgtga
     3721 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3781 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3841 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3901 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3961 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4021 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4081 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4141 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4201 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4261 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4321 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4381 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4441 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4501 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4561 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4621 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4741 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4801 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4861 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4921 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4981 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5041 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5101 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5161 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5221 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5281 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5341 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5401 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5461 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5521 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5581 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5941 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6541 gagtcagtga gcgaggaagc ggaag
//
