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
COMMENT     SalI cuts in backbone at two places.
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"
     misc_feature    94..1222
                     /label=ins
     intron          1256..1828
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    2034..2155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2198..2405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2429..3139)
                     /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(3142..3704)
                     /label=cryaa
     promoter        complement(3723..3741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3774..3790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3931..4385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4412..4516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4517..5377
                     /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      5548..6136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6424..6445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6460..6490
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6498..6514
                     /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     6522..6538
                     /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 tcgatatcgc tagcatgcat gtcgacttaa ttaaggccgc
     1921 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1981 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaatcga
     2161 gggggggccc ggtacccacc gcggtggcgg cgatctctcc ccagcatgcc tgctattgtc
     2221 ttcccaatcc tcccccttgc tgtcctgccc caacccaccc cccagaatag aatgacacct
     2281 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     2341 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     2401 cacagtcgag gctgatcagc gggccgcttt acttgtacag ctcgtccatg ccgccggtgg
     2461 agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     2521 tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct
     2581 tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct
     2641 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc
     2701 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     2761 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     2821 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt
     2881 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg
     2941 acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     3001 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     3061 cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct
     3121 cctcgccctt gctcaccatg ttgtcagacc tggtaactcc ttactgtaac ttattgcacc
     3181 tgatgaacaa aagatgtgaa ctgaacgcaa gtaggaggct aatagaggct agtacagtac
     3241 aaagatcccc aaatgatgtg tgccggcctt agtggaggca gacccctggt tatatacctg
     3301 tgctgcagaa agaggggctt acaaatggat ctctctggaa tggcattatc tcatggtggg
     3361 aacagtgttg tcagcagaat cagcggggtg gactctgacc tgagaatgac agtctagcac
     3421 tgaatggctc agacaatact cttgtggcca actatcaccg gaaaataagt gcctgccaaa
     3481 acaaaccaga ccagatgaag ctacatggct gtagaacgtc agtatttaat gtgttgtggc
     3541 cacttggttc accagattta agagctaatt tttttaaata tcacagataa tcaatcgctt
     3601 tttttttttc ttgactgaaa agcttagact ggaaaattct ttgaaaaagt ttgcactaca
     3661 gaaaataaat acataacatc ctgcactgaa tgcacactat taatggtacc cagcttttgt
     3721 tccctttagt gagggttaat tgcgcgctag ggataacagg gtaatacgcg ctcactggcc
     3781 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3841 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3901 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     3961 tcgcgttaaa tttttgttaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4081 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4141 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4201 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4261 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4321 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4381 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6541 tgattacgcc aagcgcgtta gggataacag ggtaatgcgc gc
//
