LOCUS       Exported                5103 bp ds-DNA     circular SYN 04-JUN-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5103)
  AUTHORS   rvieira
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 4, 2020 from SnapGene 5.1.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5103
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     CDS             complement(1..859)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1:   1 .. 790 / #ccffcc
                        2: 791 .. 859 / #ccffcc / signal sequence
                     Cleavage site after base 790"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
                     /note="color: black; sequence: ATCGCGAGCCCATTTATACC"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
                     /note="color: black; sequence: ATGGCTCATAACACCCCTTG"
     misc_feature    1251..1276
                     /label=MCS
                     /note="color: #333333"
     protein_bind    1293..1392
                     /label=attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #c0c0c0"
     misc_feature    1403..1436
                     /label=FRT 14
                     /note="FRT14"
                     /note="color: #ff9900; direction: RIGHT"
     misc_feature    1480..1513
                     /label=FRT
                     /note="FRT"
                     /note="color: #ffff99; direction: RIGHT"
     misc_feature    1612..1641
                     /label=SD
                     /note="Predicted by www.fruitfly.org"
                     /note="color: #00ccff"
     CDS             complement(1658..2377)
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /note="color: #00ff00"
                     /translation="GSVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGI
                     KANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     misc_feature    2399..2439
                     /label=SA
                     /note="predicted using www.fruitfly.org"
                     /note="color: #00ccff"
     misc_feature    complement(2519..2552)
                     /label=FRT 14
                     /note="FRT 14 "
                     /note="color: #ff9900; direction: LEFT"
     misc_feature    complement(2603..2636)
                     /label=FRT
                     /note="FRT"
                     /note="color: #ffff99; direction: LEFT"
     protein_bind    complement(2721..2754)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     promoter        2818..2985
                     /label=3XP3
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2986..3663
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of 
                     DsRedfluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3166..3186)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
                     /note="color: black; sequence: GTACTGGAACTGGGGGGACAG"
     primer_bind     3578..3601
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
                     /note="color: black; sequence: AGCTGGACATCACCTCCCACAACG"
     polyA_signal    3784..3905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #808080"
     primer_bind     complement(3821..3840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     3875..3894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     protein_bind    complement(3912..3945)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     protein_bind    3965..4064
                     /label=attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #c0c0c0"
     misc_feature    4076..4105
                     /label=MCS
                     /note="color: #333333"
     primer_bind     complement(4104..4121)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 
                     (-21)Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(4219..4236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      complement(4390..4978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(4470..4489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttcg ctgaagcagg tggaattctt gcatgctcct
     1261 aggtatactg cagttgctat ctattacaaa gtcccaggtc agaagcggtt ttcgggagta
     1321 gtgccccaac tggggtaacc tttgagttct ctcagttggg ggcgtagggt cgccgacatg
     1381 acacaagggg ttgaagactt ctgaagttcc tattctatca gaagtatagg aacttcagcg
     1441 gctggattgt agctgctatt agcaatatga aacctcttag aagttcctat tctctagaaa
     1501 gtataggaac ttcgaattct ggcgacttgt cgcatctttc tctgagtggg taaggtcatt
     1561 gtggggtctt aagggctaaa gggtagtgat gatgaaggca tttgatgcca ttgttcaata
     1621 acttacctcc ggaacctccg cttccaccgc tacctccctt gtacagctca tccatgccca
     1681 gggtgatgcc ggcggcggtc acgaactcca gcagcaccat gtgatcgcgc ttctcgttgg
     1741 ggtccttgga cagcacgctc tgggtgctca ggtagtggtt atcgggcagc agcactgggc
     1801 cgtcgccgat gggggtgttc tgctggtagt gatcggccag ctgcacggag ccatcctcca
     1861 cattgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttatcg gcggtgatgt
     1921 acacgttgtg gctgttgaag ttgtactcca gcttgtggcc caggatgttg ccatcctcct
     1981 tgaaatcgat gcccttcagc tcgatgcggt tcaccagggt atcgccctcg aacttcacct
     2041 cggcgcgggt cttgtaggtg ccgtcatcct tgaagctgat ggtgcgctcc tgcacgtagc
     2101 cctcgggcat ggcgctcttg aagaaatcgt gctgcttcat gtgatcgggg tagcggctga
     2161 agcactgcac gccgtaggtc agggtggtca ccagggtggg ccagggcacg ggcagcttgc
     2221 cggtggtgca gatgaacttc agggtcagct tgccgttggt ggcgtcgccc tcgccctcgc
     2281 cgcgcacgct gaacttgtgg ccgttcacgt cgccatccag ctccaccagg atgggcacca
     2341 cgccggtgaa cagctcctcg cccttggaca cggatccgga tccgccgcta cctccgcttc
     2401 caccggaacc tccgccacct gcggaagaga gataaatcgg ttgattaaaa ttagttcaag
     2461 aataattagt atatttaaac atgccgggga tgggacaagt cgccatgttg gatcgactga
     2521 agttcctata cttctgatag aataggaact tcttgcctta acccagaaat tatcactgtt
     2581 attctttaga atggtgcaaa gagaagttcc tatactttct agagaatagg aacttcccgt
     2641 cttcccgcgg acatatgcac acctgcgatc gtagtgcccc aactggggta acctttgagt
     2701 tctctcagtt gggggcgtag ataacttcgt ataatgtatg ctatacgaag ttatcgtacg
     2761 ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga tccaagctta
     2821 tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta cggagcgaca
     2881 attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca aataaacaag
     2941 cgcagctgaa caagctaaac aatcggctcg aagccggtcg ccaccatggc ctcctccgag
     3001 gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
     3061 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3121 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     3181 cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg
     3241 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3301 accgtgaccc aggactcctc cctccaggac ggctccttca tctacaaggt gaagttcatc
     3361 ggcgtgaact tcccctccga cggccccgta atgcagaaga agactatggg ctgggaggcg
     3421 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca caaggccctg
     3481 aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag
     3541 cccgtgcagc tgcccggcta ctactacgtg gactccaagc tggacatcac ctcccacaac
     3601 gaggactaca ccatcgtgga gcagtacgag cgcgccgagg gccgccacca cctgttcctg
     3661 taggggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     3721 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3781 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3841 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3901 tcttaaccgg tataacttcg tataatgtat gctatacgaa gttatagaag agcactccgt
     3961 cttcaacccc ttgtgtcatg tcggcgaccc tacgccccca actgagagaa ctcaaaggtt
     4021 accccagttg gggcactact cccgaaaacc gcttctgacc tgggaagctt gttccatccc
     4081 actcgagtac gtaggtacct cagactggcc gtcgttttac aacacagaaa gagtttgtag
     4141 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
     4201 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4801 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
     5041 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgct
     5101 ttt
//
