LOCUS       pcDNA5-FRT-TO-eG        6334 bp DNA     circular SYN 22-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5-FRT-TO-eGFP-NFATC2IP SLD1+SLD2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6334)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2024 from SnapGene Viewer 6.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6334
                     /note="/dnas_title=pcDNA5-FRT-TO-eGFP-NFATC2IP delM 
                     SLD1+SLD2"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #a6acb3"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="color: #ffffff"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tetracycline repressor TetR binding site
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="/dnas_title=tetO"
                     /note="color: #31849b"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tetracycline repressor TetR binding site
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="/dnas_title=tetO"
                     /note="color: #31849b"
     CDS             990..1706
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=enhanced GFP
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /note="/dnas_title=enhanced GFP"
                     /note="color: #993366"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1716..2225
                     /codon_start=1
                     /label=NFATC2IP delMet SLD1+SLD2 (250-418)
                     /note="NFATC2IP delMet SLD1+SLD2 (250-418)"
                     /note="color: #993366"
                     /translation="VLVEGPTLPETPRLFPLKIRCRADLVRLPLRMSEPLQSVVDHMAT
                     HLGVSPSRILLLFGETELSPTATPRTLKLGVADIIDCVVLTSSPEATETSQQLQLRVQG
                     KEKHQTLEVSLSRDSPLKTLMSHYEEAMGLSGRKLSFFFDGTKLSGRELPADLGMESGD
                     LIEVWG"
     polyA_signal    2292..2516
                     /label=bGH poly(A) signal
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    2800..2847
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /note="color: #31849b"
     CDS             2855..3877
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=aph(4)-Ia
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /note="/dnas_title=HygR"
                     /note="color: #993366"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGDFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4007..4128
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4177..4193)
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4201..4217
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
                     /note="color: #31849b"
     promoter        complement(4225..4255)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    4270..4291
                     /label=E. coli catabolite activator protein binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the
                     presence of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4579..5167)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/dnas_title=ori"
                     /note="color: #ffff00"
     CDS             complement(5338..6198)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /note="/dnas_title=AmpR"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6199..6303)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
                     /note="/dnas_title=bla"
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tggtgagcaa gggcgaggag ctgttcaccg
     1021 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1081 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1141 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1201 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1261 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1321 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1381 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1441 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1501 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1561 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1621 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1681 ctctcggcat ggacgagctg tacaagggcg cgccagtctt ggtggaaggg cccaccctcc
     1741 cagagacccc ccgactcttc ccactcaaaa tccgttgccg ggctgacctg gtcagattgc
     1801 ccctcaggat gtcggagccc ctgcagagtg tggtggacca catggccacc caccttgggg
     1861 tgtccccaag caggatcctt ttgctttttg gagagacaga gctatcacct actgccactc
     1921 ccaggaccct aaagctcgga gtggctgaca tcattgactg tgtggtacta acaagttctc
     1981 cagaggccac agagacgtcc caacagctcc agctccgggt gcagggaaag gagaaacacc
     2041 agacactgga agtctcactg tctcgagatt cccctctaaa gaccctcatg tcccactatg
     2101 aggaggccat gggactgtcg ggacggaagc tctccttctt ctttgatggg acaaagcttt
     2161 caggcaggga gctgccagct gacctgggca tggaatctgg ggacctcatt gaggtctggg
     2221 gctgagatat ccatcacact ggcggccgct cgagtctaga gggcccgttt aaacccgctg
     2281 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2341 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2401 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2461 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2521 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2581 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2641 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2701 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2761 ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc
     2821 ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc
     2881 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     2941 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3001 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     3061 cgcgctcccg attccggaag tgcttgacat tggggatttc agcgagagcc tgacctattg
     3121 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3181 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     3241 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3301 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3361 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3421 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3481 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3541 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     3601 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     3661 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     3721 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     3781 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     3841 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc
     3901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4141 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6301 gcgcacattt ccccgaaaag tgccacctga cgtc
//
