LOCUS       Exported                7549 bp ds-DNA     circular SYN 16-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7549)
  AUTHORS   Richard Davis
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 19, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7549
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    33..80
                     /label=FRT-F3 mutant
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
     rep_origin      347..802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1084..1188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1189..2049
                     /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      2220..2808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3096..3117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3132..3162
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3170..3186
                     /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        3242..3452
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3548..4725
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_recomb     4743..4784
                     /label=attB
                     /note="mutant attB recombination sequence for bxb1"
     protein_bind    4797..4844
                     /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        4892..4893
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_feature    4896..6552
                     /label=CAG
     CDS             6611..7309
                     /codon_start=1
                     /label=mStayGold
                     /translation="MRGSDPMVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSH
                     EGSHKGKYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQ
                     YKGDGSIHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTSDMDVSLPNEVQHIPIDDGV
                     ECTVTLQYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHI
                     IQSETLEAHL"
     polyA_signal    7321..7545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgctcgctgc aagaactctt cctcacggat ccgaagttcc tattccgaag ttcctattct
       61 tcaaatagta taggaacttc agcgctcact ggccgtcgtt ttacaaaggg cgaattcact
      121 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      181 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      241 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
      301 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
      361 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      421 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      481 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      541 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
      601 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      661 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
      721 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      781 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
      841 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      901 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      961 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     1021 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     1081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3001 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3061 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     3121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     3181 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttaa ttccctcccc
     3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttgca aagatggata
     3481 aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg
     3541 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3601 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     3661 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3721 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     3781 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     3841 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     3901 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     3961 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     4021 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     4081 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     4141 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     4201 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     4261 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     4321 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     4381 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     4441 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     4501 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     4561 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     4621 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     4681 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tcgccaccat
     4741 ggccggcttg tcgacgacgg cggactccgt cgtcaggatc atccggatcc ggatccgaag
     4801 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcatgata ttcggcaagc
     4861 aggcatcgac tagttaatta agtacttact actagttatt aatagtaatc aattacgggg
     4921 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4981 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     5041 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5101 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5161 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5221 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     5281 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     5341 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccrgg sggggsgggg
     5401 sggggsgrgg ggsggggsgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
     5461 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
     5521 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
     5581 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     5641 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     5701 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     5761 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
     5821 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     5881 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
     5941 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
     6001 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     6061 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gctggtgggg
     6121 gtgccgggcg gggcggggcc gcctcgggcc ggggrgggct cgggggaggg gcgcggcggc
     6181 ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa
     6241 tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg
     6301 aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag
     6361 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag
     6421 cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
     6481 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
     6541 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
     6601 aatggccacc atgcggggtt ctgatccgat ggtgtctaca ggcgaggagc tgtttaccgg
     6661 cgtggtgccc ttcaagttcc agctgaaggg caccatcaac ggcaagagct tcaccgtgga
     6721 aggcgagggc gagggcaata gccacgaggg cagccacaaa ggcaagtacg tgtgcaccag
     6781 cggcaaactg ccaatgtctt gggccgccct gggaactagc ttcggctatg gcatgaagta
     6841 ctacaccaag taccccagcg gcctgaagaa ctggttccac gaggtgatgc ccgagggctt
     6901 cacctacgac agacacatcc agtacaaggg cgacggcagc atccacgcca agcaccagca
     6961 cttcatgaag aacggcacct accacaacat cgtggagttc accggccagg acttcaagga
     7021 gaacagcccc gtgctgacca gcgacatgga cgtgagcctg cccaacgagg tgcagcacat
     7081 ccccatagat gacggcgtgg agtgcacagt gaccctgcag taccctctgc tgagcgacga
     7141 gagcaagtgc gtggaggcct accagaacac catcatcaag cccctgcaca atcagccagc
     7201 ccccgatgtg ccattccact ggatcagaaa gcagtacacc cagagcaagg acgacaccga
     7261 ggagagggac cacatcatcc agagcgagac actggaggcc cacctgtaac aacgttgcac
     7321 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     7381 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     7441 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     7501 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcgct
//
