LOCUS       Exported                8178 bp DNA     circular SYN 20-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8178)
  AUTHORS   Tyler Kirby
  TITLE     Direct Submission
  JOURNAL   Exported May 31, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    224..305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        434..1611
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          665..1602
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             1669..2415
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    2434..3020
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3021..3824
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3838..4062
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4387..4975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5146..6006)
                     /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"
     promoter        complement(6007..6111)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    6490..6760
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     intron          7062..7194
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             7244..7960
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQNNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
       61 atcaatcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
      121 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
      181 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
      241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      301 actgcattct agttgtggtt tgtccaaact catcaatgct cgacgataag ctttgcaaag
      361 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
      421 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
      481 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt
      541 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      601 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      661 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
      721 cgtgccttga attacttcca ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
      781 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
      841 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
      901 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
      961 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     1021 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     1081 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     1141 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     1201 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     1261 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     1321 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     1381 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     1441 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     1501 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     1561 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaatttcg
     1621 acatttaaat ttaattaatc tcgacggtat cggttgggat ctgccaccat gtccagactg
     1681 gacaagagca aagtcataaa ctctgctctg gaattactca atgaagtcgg tatcgaaggc
     1741 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     1801 gtgaagaaca agcgggccct gctcgatgcc ctggcaatcg agatgctgga caggcatcat
     1861 acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     1921 tcattccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     1981 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     2041 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     2101 gtattggagg atcaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     2161 tctatgcccc cacttctgag acaagcaatt gagctgttcg accatcaggg agccgaacct
     2221 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     2281 agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     2341 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     2401 atgctccccg gctaacccaa acccgctgat ccgcccctct ccctcccccc cccctaacgt
     2461 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2521 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2581 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2641 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2701 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2761 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     2821 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     2881 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     2941 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     3001 atgataatat ggccacaacc atgggatcgg ccattgaaca agatggattg cacgcaggtt
     3061 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3121 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3181 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3241 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3301 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3361 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3421 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3481 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3541 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3601 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3661 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3721 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3781 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgatgatca gcctcgactg
     3841 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3901 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3961 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4021 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcatcgatg ctagttaaaa
     4081 gttttgttac tttatagaag aaattttgag tttttgtttt tttttaataa ataaataaac
     4141 ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata ataaaactta
     4201 atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca catgcgtcaa
     4261 ttttacgcat gattatcttt aacgtacgtc acaatatgat tatctttcta gggttaatct
     4321 agtatacgcg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4801 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6121 ccgaaaagtg ccaccgctag attaacccta gaaagatagt ctgcgtaaaa ttgacgcatg
     6181 cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg tgcatttagg
     6241 acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg taagtgtcac
     6301 tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct tttacgtgac
     6361 ttttaagatt taactcatac gataattata ttgttatttc atgttctact tacgtgataa
     6421 cttattatat atatattttc ttgttataga tatcgggtca gcttggccta gcgctgtcga
     6481 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     6541 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     6601 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     6661 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     6721 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     6781 tacccgggtc gagtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga
     6841 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     6901 accgatccag cctccgcgga atttgattag ctttattgcg gtagtttatc acagttaaat
     6961 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gtgactctct
     7021 taaggtagcc ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
     7081 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     7141 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
     7201 actcccagtt caattacagc tcttaaggct agaggatccc accatggtga gcaagggcga
     7261 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     7321 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     7381 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     7441 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     7501 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     7561 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     7621 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     7681 caacagccac aacgtctata tcatggccga caagcagaat aacggcatca aggtgaactt
     7741 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     7801 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     7861 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     7921 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctagctc
     7981 ccagcctaga cgatccttcc tctcaagggt gatcagggca gcgctaccgt tgcagctgct
     8041 tctgctgctg ctgctgctcc tggcctgcct gctacctgcc tctgaagatg actacagctg
     8101 cacccaggcc aacaactttg cccgatcctt ctaccccatg ctgcggtaca ccaacgggcc
     8161 acctcccacc taggaatt
//
