LOCUS       Exported                8121 bp ds-DNA     circular SYN 17-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8121)
  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..8121
                     /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        1888..3544
                     /label=CAG
     regulatory      3605..3610
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3614..4312
                     /codon_start=1
                     /label=mStayGold
                     /translation="MRGSDPMVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSH
                     EGSHKGKYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQ
                     YKGDGSIHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGV
                     ECTVTLQYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHI
                     IQSETLEAHL"
     misc_feature    4313..4901
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4954..5178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5210..5242
                     /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      5479..5934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6216..6320
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6321..7181
                     /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      7352..7940
                     /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 taagtactta ctactagtta ttaatagtaa tcaattacgg ggtcattagt
     1921 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1981 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2041 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2101 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     2161 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     2221 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     2281 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     2341 tgcagcgatg ggggcggggg gggggggggg gcgcgcgccr ggsggggsgg ggsggggsgr
     2401 ggggsggggs ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
     2461 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
     2521 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
     2581 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
     2641 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
     2701 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
     2761 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
     2821 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
     2881 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
     2941 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
     3001 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     3061 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
     3121 cggggcgggg ccgcctcggg ccggggrggg ctcgggggag gggcgcggcg gcccccggag
     3181 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     3241 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     3301 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     3361 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     3421 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     3481 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     3541 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aaaatggcca
     3601 ccatgccacc atgatgcggg gttctgatcc gatggtgtct acaggcgagg agctgtttac
     3661 cggcgtggtg cccttcaagt tccagctgaa gggcaccatc aacggcaaga gcttcaccgt
     3721 ggaaggcgag ggcgagggca atagccacga gggcagccac aaaggcaagt acgtgtgcac
     3781 cagcggcaaa ctgccaatgt cttgggccgc cctgggaact agcttcggct atggcatgaa
     3841 gtactacacc aagtacccca gcggcctgaa gaactggttc cacgaggtga tgcccgaggg
     3901 cttcacctac gacagacaca tccagtacaa gggcgacggc agcatccacg ccaagcacca
     3961 gcacttcatg aagaacggca cctaccacaa catcgtggag ttcaccggcc aggacttcaa
     4021 ggagaacagc cccgtgctga ccggcgacat ggacgtgagc ctgcccaacg aggtgcagca
     4081 catccccata gatgacggcg tggagtgcac agtgaccctg cagtaccctc tgctgagcga
     4141 cgagagcaag tgcgtggagg cctaccagaa caccatcatc aagcccctgc acaatcagcc
     4201 agcccccgat gtgccattcc actggatcag aaagcagtac acccagagca aggacgacac
     4261 cgaggagagg gaccacatca tccagagcga gacactggag gcccacctgt aaaatcaacc
     4321 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4381 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4441 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4501 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4561 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4621 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4681 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4741 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4801 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4861 ccctcagacg agtcggatct ccctttgggc cgcctccccg ccctttgggc cgcctccccg
     4921 cgctagtgct ggggatgcgt aacaacgttg cacctgtgcc ttctagttgc cagccatctg
     4981 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     5041 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     5101 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     5161 atgcggtggg ctctatggcg ctcgctcgct gcaagaactc ttcctcacga taacttcgta
     5221 taggagactt tatacgaagt taagcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5281 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5341 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5401 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5461 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5521 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5821 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5881 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5941 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     6001 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     6061 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6121 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6181 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6241 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6301 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6421 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6481 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6541 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6601 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6721 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6841 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6901 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7141 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7981 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8041 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8101 cagtgagcga ggaagcggaa g
//
