LOCUS       Exported                6582 bp ds-DNA     circular SYN 18-MAR-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   Sumeet Pal Singh
  TITLE     Direct Submission
  JOURNAL   Exported Apr 3, 2018 from SnapGene Viewer 4.1.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6582
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    37..61
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        94..1222
                     /label=ins
     intron          1256..1828
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    2033..2154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2197..2404
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2428..3138)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    complement(3141..3703)
                     /label=cryaa
     promoter        complement(3722..3740)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3773..3789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3930..4384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4411..4515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4516..5376
                     /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      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6423..6444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6459..6489
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6497..6513
                     /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)."
     primer_bind     6521..6537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctcatcacaa gtttgtacaa aaaagcaggc
       61 tccgcggccg cccccttcac cacgcgtaag ctttgtttgt accataataa gctcattata
      121 taatatttaa aatgatactg catcattaat tgatatgttt tctagttatt tttaaatatt
      181 agcaactaaa ttaggaaatt atccatagta aatttaatgt aatattattt gttatattta
      241 acttcagccc acagtctagt ttagttgttg gtcccttatg agaaaaaatt tgggcacctc
      301 tgctctagtg cttgtaaaca tttttgagta tctatagttg aacatgaaag catttttttt
      361 tttttttttt taagaaaatg aagaaagcta aaaacctgta tccactgact tccattaaaa
      421 aaagcaaata ctagggatgt ctatggtgtc ttcttttgtg ttcaacaaat aagcagaaag
      481 taactgctga cagaacatta cattttgtgt gaatcccctt atattgtgtt aaaaactgta
      541 gagaaatcat ctaataataa gaaattatca agaaataaac agaaaatgat ctaatagtat
      601 actataaaat ctttcaaaat ccaggccact accatatgac taaaaccaca catattatgc
      661 tttaaaaaaa gtcacaatga agacgtcaca aacgttcctg aagactaatg actcgtctaa
      721 ccccatctta ccttctgctg tttcactcaa agaaaagcca ccagctgtct ctgtgatctg
      781 ctcattctgt gttaacttgt gttctcaaca attaacaatt aaccaaaagc ctctggagct
      841 gcgtgagcat cacgatgtgt ttgtcttctg tgagcgtggc agtgtccacg aagtcaatgt
      901 gaattggttg gccaaacaca gctaatgtgg catgtgtgat tggacagcca cttaaaagtg
      961 ttaaagtgtg tttacgcaca ggattggttt tccgctgaat caggcaaaac cagcagaacc
     1021 gatctaaact cgagtataaa agtgagtagg agatgctatt cgtcccaaaa catctccacc
     1081 accatctcca ccattcctcg cctctgcttc gagaacaggt gagtgtcgag cgggatggta
     1141 aatctacaga gaatgcggag tgtagcttgt gtacatgttt ttgattaaca gagattgtat
     1201 gtgtgtgttt gtgtcagtgt gaaagggtgg gcgcgccgat cctgagaact tcagggtgag
     1261 tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt tatatggagg
     1321 gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc accatggacc
     1381 ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt ctcctcttat
     1441 tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt aacgaatttt
     1501 taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac ttttttttca
     1561 aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa ttgttataat
     1621 taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa atattcttat
     1681 tggtagaaac aactacaccc tggtcatcat cctgcctttc tctttatggt tacaatgata
     1741 tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct ctgctaacca
     1801 tgttcatgcc ttcttctctt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc
     1861 tcatcatttt ggcaaagaat tactagtgga tccgatatcg aattcttaat taaggccgcg
     1921 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1981 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaatcgag
     2161 ggggggcccg gtacccaccg cggtggcggc gatctctccc cagcatgcct gctattgtct
     2221 tcccaatcct cccccttgct gtcctgcccc aacccacccc ccagaataga atgacaccta
     2281 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2341 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2401 acagtcgagg ctgatcagcg ggccgcttta cttgtacagc tcgtccatgc cgccggtgga
     2461 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     2521 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     2581 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     2641 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     2701 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     2761 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     2821 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     2881 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     2941 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     3001 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     3061 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     3121 ctcgcccttg ctcaccatgt tgtcagacct ggtaactcct tactgtaact tattgcacct
     3181 gatgaacaaa agatgtgaac tgaacgcaag taggaggcta atagaggcta gtacagtaca
     3241 aagatcccca aatgatgtgt gccggcctta gtggaggcag acccctggtt atatacctgt
     3301 gctgcagaaa gaggggctta caaatggatc tctctggaat ggcattatct catggtggga
     3361 acagtgttgt cagcagaatc agcggggtgg actctgacct gagaatgaca gtctagcact
     3421 gaatggctca gacaatactc ttgtggccaa ctatcaccgg aaaataagtg cctgccaaaa
     3481 caaaccagac cagatgaagc tacatggctg tagaacgtca gtatttaatg tgttgtggcc
     3541 acttggttca ccagatttaa gagctaattt ttttaaatat cacagataat caatcgcttt
     3601 ttttttttct tgactgaaaa gcttagactg gaaaattctt tgaaaaagtt tgcactacag
     3661 aaaataaata cataacatcc tgcactgaat gcacactatt aatggtaccc agcttttgtt
     3721 ccctttagtg agggttaatt gcgcgctagg gataacaggg taatacgcgc tcactggccg
     3781 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3841 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3901 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt
     3961 cgcgttaaat ttttgttaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4021 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4081 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4141 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4201 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4261 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4321 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4381 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5221 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6301 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6361 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6421 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6481 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6541 gattacgcca agcgcgttag ggataacagg gtaatgcgcg cg
//
