LOCUS       Exported                6598 bp ds-DNA     circular SYN 15-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-Cre_T2A_mRuby2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6598)
  AUTHORS   Pallab
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 17, 2018 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6598
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /note="CMV enhancer"
                     /note="CMV enhancer;human cytomegalovirus immediate early 
                     enhancer;color: #ffffff"
     promoter        386..661
                     /note="chicken beta-actin promoter"
                     /note="chicken beta-actin promoter;color: #ffffff; 
                     direction: RIGHT"
     intron          662..1670
                     /note="chimeric intron"
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    662..1625
                     /note="Chicken beta actin intron"
                     /note="Chicken beta actin intron;color: #ff9900; direction:
                     RIGHT"
     misc_feature    1725..1730
                     /note="Kozak"
                     /note="Kozak;color: #a6acb3"
     CDS             1731..2759
                     /codon_start=1
                     /product="site-specific recombinase"
                     /note="site-specific recombinase"
                     /note="Cre;Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP 
                     sequences.;color: #993366"
                     /protein_id=""
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    2760..2768
                     /note="GSG"
                     /note="GSG;to increase cleavage efficiency of 2A 
                     Peptide;color: #cc99ff; direction: RIGHT"
     CDS             2769..2822
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /note="T2A;Eukaryotic ribosomes fail to insert a peptide 
                     bond between the Gly and Pro residues, yielding separate 
                     polypeptides.;color: #cc99b2 Cleavage site after base 2819"
                     /protein_id=""
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2823..3533
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /note="monomeric red fluorescent protein derived from"
                     /note="mRuby2;color: #ff0000"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             3540
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /note="Myc;color: #cc99b2"
                     /translation=""
     polyA_site      3687..3742
                     /note="beta-globin poly(A) signal"
                     /note="beta-globin poly(A) signal;rabbit beta-globin 
                     polyadenylation signal;color: #a6acb3"
     primer_bind     complement(4101..4117)
                     /note="M13 rev"
                     /note="M13 rev;common sequencing primer, one of multiple 
                     similar variants;color: #a020f0; direction: LEFT"
     protein_bind    4125..4141
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac repressor encoded by lacI lac operator"
                     /note="lac operator;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).;color: 
                     #31849b"
     promoter        complement(4149..4179)
                     /note="lac promoter"
                     /note="lac promoter;promoter for the E. coli lac 
                     operon;This reverse directional feature has 3 segments: 1: 
                     4149 .. 4155 / #ffffff / -10 2: 4156 .. 4173 / #ffffff 3: 
                     4174 .. 4179 / #ffffff / -35"
     protein_bind    4194..4215
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="E. coli catabolite activator protein CAP bindin"
                     /note="CAP binding site;CAP binding activates transcription
                     in the presence of cAMP.;color: #31849b"
     promoter        4274..4469
                     /note="SV40 promoter"
                     /note="SV40 promoter;SV40 early promoter;color: #ffffff; 
                     direction: RIGHT"
     rep_origin      4320..4455
                     /note="SV40 ori"
                     /note="SV40 ori;SV40 origin of replication;color: #ffff00"
     polyA_signal    4475..4609
                     /note="SV40 poly(A) signal"
                     /note="SV40 poly(A) signal;SV40 polyadenylation 
                     signal;color: #a6acb3"
     rep_origin      complement(4848..5436)
                     /direction=LEFT
                     /note="ori"
                     /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication;color: #ffff00"
     CDS             complement(5607..6467)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="bla"
                     /note="AmpR;confers resistance to ampicillin, 
                     carbenicillin, and related antibiotics;This feature has 2 
                     segments: 1: 5607 .. 6398 / #ccffcc 2: 6399 .. 6467 / 
                     #ccffcc / signal sequence Cleavage site after base 6398"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6468..6572)
                     /gene="bla"
                     /note="bla AmpR promoter"
                     /note="AmpR promoter;color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccacc atggccaatt
     1741 tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc
     1801 gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa
     1861 tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt
     1921 ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg
     1981 cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc
     2041 tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa
     2101 acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc
     2161 aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc
     2221 tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag
     2281 atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg
     2341 ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat
     2401 ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa
     2461 aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga
     2521 tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt cagagatacc
     2581 tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag
     2641 tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact
     2701 atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa gatggcgatg
     2761 ggagtggaga gggcagggga agtcttctaa catgcgggga cgtggaggaa aatcccggcc
     2821 ccatggtgtc taagggcgaa gagctgatca aggaaaatat gcgtatgaag gtggtcatgg
     2881 aaggttcggt caacggccac caattcaaat gcacaggtga aggagaaggc aatccgtaca
     2941 tgggaactca aaccatgagg atcaaagtca tcgagggagg acccctgcca tttgcctttg
     3001 acattcttgc cacgtcgttc atgtatggca gccgtacttt tatcaagtac ccgaaaggca
     3061 ttcctgattt ctttaaacag tcctttcctg agggttttac ttgggaaaga gttacgagat
     3121 acgaagatgg tggagtcgtc accgtcatgc aggacaccag ccttgaggat ggctgtctcg
     3181 tttaccacgt ccaagtcaga ggggtaaact ttccctccaa tggtcccgtg atgcagaaga
     3241 agaccaaggg ttgggagcct aatacagaga tgatgtatcc agcagatggt ggtctgaggg
     3301 gatacactca tatggcactg aaagttgatg gtggtggcca tctgtcttgc tctttcgtaa
     3361 caacttacag gtcaaaaaag accgtcggga acatcaagat gcccggtatc catgccgttg
     3421 atcaccgcct ggaaaggtta gaggaaagtg acaatgaaat gttcgtagta caacgcgaac
     3481 acgcagttgc caagttcgcc gggcttggtg gtgggatgga cgagctgtac aagtgagcgg
     3541 ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     3601 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     3661 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     3721 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     3781 tcagaatgag tatttggttt agagtttggc aacatatgcc atatgctggc tgccatgaac
     3841 aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     3901 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     3961 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     4021 cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc gacctgcagc
     4081 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4141 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4201 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4261 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4321 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4381 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4441 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     4501 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4561 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     4621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5041 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5281 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6481 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6541 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
