LOCUS       Exported                8235 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 8235)
  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..8235
                     /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    1808..1855
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1894..3560
                     /label=CAG promoter
     regulatory      3632..3637
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    3649..3670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3685..3715
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3723..3739
                     /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)."
     CDS             3759..4082
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     region          4083..4120
                     /label=terminator
     misc_feature    4148..4736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4775..5250
                     /label=Rabbit bGlobulin polyAd signal
                     /note="Rabbit bGlobulin polyAd signal"
     protein_bind    5279..5326
                     /label=FRT-F3 mutant
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
     rep_origin      5593..6048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6330..6434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6435..7295
                     /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      7466..8054
                     /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 gcggactccg tcgtcaggat catccggatc
     1801 cggatccgaa gttcctattc cgaagttcct attctctaga aagtatagga acttcatgat
     1861 attcggcaag caggcatcga ctagttaatt aaggacattg attattgact agttattaat
     1921 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     1981 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2041 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggact
     2101 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2161 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2221 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgggtcgag
     2281 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     2341 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggcgcgc
     2401 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     2461 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     2521 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgttgcctt cgccccgtgc
     2581 cccgctccgc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     2641 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     2701 gctcgtttct tttctgtggc tgcgtgaaag ccttaaaggg ctccgggagg gccctttgtg
     2761 cgggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg
     2821 gcccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     2881 cgtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg ggctgcgagg
     2941 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgg
     3001 cggtcgggct gtaacccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     3061 tcgggtgcgg ggctccgtgc ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     3121 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     3181 gggcgcggcg gccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     3241 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tggcggagcc
     3301 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc gggcgaagcg gtgcggcgcc
     3361 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     3421 catctccagc ctcggggctg ccgcaggggg acggctgcct tcggggggga cggggcaggg
     3481 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     3541 ccttcttctt tttcctacag ctcctgggca acgtgctggt tgttgtgctg tctcatcatt
     3601 ttggcaaaga attcctcgag atccattgtg ggccaccatg gcggccgcta atgtgagtta
     3661 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     3721 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     3781 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat tcactggccg
     3841 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3901 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3961 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     4021 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4081 agcccagccc gcctaatgag cgggcttttt tttgaacaaa ttaagcctat aacgcggaag
     4141 cggccgcaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4201 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4261 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4321 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4381 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4441 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4501 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4561 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4621 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4681 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgccctt
     4741 tgggccgcct ccccgcgcta gtgctgggga tgcgccaatg ccctggctca caaataccac
     4801 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4861 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4921 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4981 gtttagagtt tggcaacata tgccatatgc tggctgccat gaacaaaggt ggctataaag
     5041 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     5101 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     5161 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     5221 catagctgtc cctcttctct tatgaagatc cgctgcaaga actcttcctc acggatccga
     5281 agttcctatt ccgaagttcc tattcttcaa atagtatagg aacttcagcg ctcactggcc
     5341 gtcgttttac aaagggcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5401 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5461 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5521 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5581 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5641 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5881 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5941 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6001 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     6061 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6121 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6181 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6241 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6301 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6481 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6541 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6601 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6661 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6721 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6781 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6841 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6901 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6961 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7021 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7081 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7141 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7201 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7261 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7321 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     7621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8101 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8161 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8221 gcgaggaagc ggaag
//
