LOCUS       Exported                9182 bp ds-DNA     circular SYN 13-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9182)
  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..9182
                     /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)."
     misc_feature    1900..3493
                     /label=pCAG
     regulatory      3555..3562
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3572..4267
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             4295..4990
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             5018..5713
                     /codon_start=1
                     /product="improved green fluorescent protein from 
                     Pontellina plumata (Evdokimov et al., 2006)"
                     /label=TurboGFP
                     /note="mammalian codon-optimized"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     CDS             5732..5752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5756..5776
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5780..5800
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5816..5938
                     /label=PEST
     polyA_signal    6072..6193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6226..6273
                     /label=FRT-F3 mutant
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
     rep_origin      6540..6995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7277..7381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7382..8242
                     /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      8413..9001
                     /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 aagtttccgc gttacataac ttacggtaaa
     1921 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     1981 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2041 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2101 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2161 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
     2221 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
     2281 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg
     2341 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     2401 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     2461 agcgaagcgc gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg
     2521 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
     2581 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt
     2641 ttctgtggct gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc gggggggagc
     2701 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc
     2761 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga
     2821 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc
     2881 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc ggtcgggctg
     2941 taaccccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg
     3001 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg
     3061 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg
     3121 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     3181 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     3241 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     3301 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     3361 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     3421 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
     3481 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
     3541 agaattccga tcgcgccacc atcctggacc tatggagagc gacgagagcg gcctgcccgc
     3601 catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg
     3661 cggcggagag ggcacccccg agcagggccg catgaccaac aagatgaaga gcaccaaagg
     3721 cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt
     3781 cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta
     3841 caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta
     3901 ccgctacgag gccggccgcg tgatcggcga cttcaaggtg atgggcaccg gcttccccga
     3961 ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca
     4021 ccccatgggc gataacgatc tggatggcag cttcacccgc accttcagcc tgcgcgacgg
     4081 cggctactac agctccgtgg tggacagcca catgcacttc aagagcgcca tccaccccag
     4141 catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagg atcacagcaa
     4201 caccgagctg ggcatcgtgg agtaccagca cgccttcaag accccggatg cagatgccgg
     4261 tgaagaactc agatccactg gaggcggagg tggaatggag agcgacgaga gcggcctgcc
     4321 cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt
     4381 gggcggcgga gagggcaccc ccgagcaggg ccgcatgacc aacaagatga agagcaccaa
     4441 aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca
     4501 cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg
     4561 ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag
     4621 ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtgatgggca ccggcttccc
     4681 cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct
     4741 gcaccccatg ggcgataacg atctggatgg cagcttcacc cgcaccttca gcctgcgcga
     4801 cggcggctac tacagctccg tggtggacag ccacatgcac ttcaagagcg ccatccaccc
     4861 cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg aggatcacag
     4921 caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccgg atgcagatgc
     4981 cggtgaagaa ctcagatcca ctggaggcgg aggtggaatg gagagcgacg agagcggcct
     5041 gcccgccatg gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct
     5101 ggtgggcggc ggagagggca cccccgagca gggccgcatg accaacaaga tgaagagcac
     5161 caaaggcgcc ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta
     5221 ccacttcggc acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg
     5281 cggctacacc aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt
     5341 cagctaccgc tacgaggccg gccgcgtgat cggcgacttc aaggtgatgg gcaccggctt
     5401 ccccgaggac agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca
     5461 cctgcacccc atgggcgata acgatctgga tggcagcttc acccgcacct tcagcctgcg
     5521 cgacggcggc tactacagct ccgtggtgga cagccacatg cacttcaaga gcgccatcca
     5581 ccccagcatc ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggaggatca
     5641 cagcaacacc gagctgggca tcgtggagta ccagcacgcc ttcaagaccc cggatgcaga
     5701 tgccggtgaa gaactcagat ctcgagctga tccaaaaaag aagagaaagg tagatccaaa
     5761 aaagaagaga aaggtagatc caaaaaagaa gagaaaggta ggatccaccg gatctagcca
     5821 tggcttcccg ccggaggtgg aggagcagga tgatggcacg ctgcccatgt cttgtgccca
     5881 ggagagcggg atggaccgtc accctgcagc ctgtgcttct gctaggatca atgtgtagtg
     5941 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     6001 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     6061 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6121 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6181 tcaatgtatc ttaaggcggc tgcaagaact cttcctcacg gatccgaagt tcctattccg
     6241 aagttcctat tcttcaaata gtataggaac ttcagcgctc actggccgtc gttttacaaa
     6301 gggcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6361 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6421 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     6481 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     6541 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6601 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6661 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6721 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     6781 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6841 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     6901 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6961 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     7021 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     7081 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7141 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     7201 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     7261 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7321 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7381 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7441 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7501 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7561 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7621 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7681 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7741 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7801 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7861 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7921 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7981 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     8041 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     8101 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8161 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8221 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8281 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8341 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8401 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8461 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8521 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     8581 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8641 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8701 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8761 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8821 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8881 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8941 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     9001 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     9061 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     9121 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     9181 ag
//
