LOCUS       Exported                7885 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCalN-ddGFP-A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7885)
  AUTHORS   Alford SC, Ding Y, Simmen T, Campbell RE
  TITLE     Dimerization-dependent green and yellow fluorescent proteins.
  JOURNAL   ACS Synth Biol. 2012 Dec 21;1(12):569-75. doi: 10.1021/sb300050j. 
            Epub 2012 Aug 14.
  PUBMED    23656278
REFERENCE   2  (bases 1 to 7885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7885
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      906..915
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2875..2893)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3143..3162
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3337..3360
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3476..3493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3482..3706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3752..4180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3839..3858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4049..4070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4189..4209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4194..4523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4374..4509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4436..4455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4590..5384
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4644..4663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5254..5273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5558..5679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5595..5614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5649..5668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5728..5744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5728..5744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5741..5763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5752..5768
                     /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)."
     promoter        complement(5776..5806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5821..5842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5959..5976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6130..6718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6210..6229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6889..7749)
                     /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"
     primer_bind     7512..7531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7750..7854)
                     /gene="bla"
                     /label=AmpR promoter
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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtgccgccac catggcctcg agaatggaag ggaagtggtt gctgtgtatg ttactggtgc
      961 ttggaactgc tattgttgag gctcatgatg gacatgatga tgatgtgatt gatattgagg
     1021 atgaccttga cgatgtcatt gaagaggtag aagactcaaa accagatacc actgctcctc
     1081 cttcatctcc caaggttact tacaaagctc cagttccaac aggggaagta tattttgctg
     1141 attcttttga cagaggaact ctgtcagggt ggattttatc caaagccaag aaagacgata
     1201 ccgatgatga aattgccaaa tatgatggaa agtgggaggt agaggaaatg aaggagtcaa
     1261 agcttccagg tgataaagga cttgtgttga tgtctcgggc caagcatcat gccatctctg
     1321 ctaaactgaa caagcccttc ctgtttgaca ccaagcctct cattgttcag tatgaggtta
     1381 atttccaaaa tggaatagaa tgtggtggtg cctatgtgaa actgctttct aaaacaccag
     1441 aactcaacct ggatcagttc catgacaaga ccccttatac gattatgttt ggtccagata
     1501 aatgtggaga ggactataaa ctgcacttca tcttccgaca caaaaacccc aaaacgggta
     1561 tctatgaaga aaaacatgct aagaggccag atgcagatct gaagacctat tttactgata
     1621 agaaaacaca tctttacaca ctaatcttga atccagataa tagttttgaa atactggttg
     1681 accaatctgt ggtgaatagt ggaaatctgc tcaatgacat gactcctcct gtaaatcctt
     1741 cacgtgaaat tgaggaccca gaagaccgga agcccgagga ttgggatgaa agaccaaaaa
     1801 tcccagatcc agaagctgtc aagccagatg actgggatga agatgcccct gctaagattc
     1861 cagatgaaga ggccacaaaa cccgaaggct ggttagatga tgagcctgag tacgtacctg
     1921 atccagacgc agagaaacct gaggattggg atgaagacat ggatggagaa tgggaggctc
     1981 ctcagattgc caaccctaga tgtgagtcag ctcctggatg tggtgtctgg cagcgacctg
     2041 tgattgacaa ccccaattat aaaggcaaat ggaagcctcc tatgattgac aatcccagtt
     2101 accagggaat ctggaaaccc aggaaaatac caaatccaga tttctttgaa gatctggaac
     2161 ctttcagaat gactcctttt agtgctattg gtttggagct gtggtccatg acctctgaca
     2221 ttttttttga caactttatc atttgtgctg atcgaagaat agttgatgat tgggccaatg
     2281 atggatgggg cctgaagaaa gctgctgatg gggctgctga gccaggcgtt gtggggcaga
     2341 tgaacgaggc agctgaagag cgcccgtggc tgtgggtagt ctatattcta actgtagccc
     2401 ttcctgtgtt cctggttatc ctcttctgct gttctggaaa gaaacagacc agtggtatgg
     2461 agtataagaa aactgatgca cctcaaccgg atgtgaagga agaggaagaa gagaaggaag
     2521 aggaaaagga caagggagat gaggaggagg aaggagaaga gaaacttgaa gagaaacaga
     2581 aaagtgatgc tgaagaagat ggtggcactg tcagtcaaga ggaggaagac agaaaaccta
     2641 aagcagagga ggatgaaatt ttgaacagat caccaagaaa cagaaagcca cgaagagagg
     2701 ggggtgcgtc aggcggcagc ggatctggac cggtcggtac catggtgagc aagagcgagg
     2761 aggtcatcaa agagttcatg cgcttcaagg tgcgcttgga gggctccatg aacggccacg
     2821 agttcgagat cgagggcgag ggcgagggcc gcccttacga gggcacccag accgccaagc
     2881 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc cccctgatca
     2941 tgtacggctc caagatgtac gtgaagcacc ccgccgacgt ccccgattac atgaagctgt
     3001 ccttccccga gggcttcaag tgggagcgcg tgatgcactt cgaggacggc ggtctggtga
     3061 ccgcgacaca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     3121 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccttgggc tgggattatg
     3181 ccaccgagcg cctgtacccc gaagaaggcg tgctgaaggg cgagctcctg gggcgcctga
     3241 agctgaagga cggcggcctc aacctggtgg agtccaagac catctacatg gccaagaagc
     3301 ccgtgcaact gcccggctac tacttcgtgg acaccaagct ggacatcacc tcccacaacg
     3361 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctggggcatg
     3421 gtacaggtag cacaggcagc ggtagctcct aaaagcttgg taaaccgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3721 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3961 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4021 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4081 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4141 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4201 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4561 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4621 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4681 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4741 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4801 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4861 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4921 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4981 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5041 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5101 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5161 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5221 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5281 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5341 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5401 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5461 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5521 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5701 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctatagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7861 tccccgaaaa gtgccacctg acgtc
//
