LOCUS       Exported                4839 bp ds-DNA     circular SYN 06-MAR-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Mar 6, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4839
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     594..610
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        620..638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    647..651
                     /label=MCS
                     /note="color: #808080; direction: RIGHT"
     misc_feature    652..672
                     
                     /note="color: #0000ff; direction: RIGHT"
     CDS             657..1592
                     /codon_start=1
                     /label=BamHI-Linker-mAID-mCherry2
                     /note="color: #ff0000"
                     /translation="GSGAGAKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKS
                     SGGPEAAAFVKVSMDGAPYLRKIDLRMYKASMVSKGEEDNMAIIKEFMRFKVHMEGSVN
                     GHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDY
                     LKLSFPEGFNWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTM
                     GWEASTERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLD
                     ILSHNEDYTIVEQYERAEGRHSTGGMDELYK"
     CDS             657..674
                     /codon_start=1
                     /label=Linker
                     /note="note=""direct strand 
                     translation""translation=""GAGA"""
                     /note="color: #ff6666"
                     /translation="GSGAGA"
     CDS             675..878
                     /codon_start=1
                     /label=mAID
                     /note="note=""direct strand 
                     translation""translation=""KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNV
                     MVSCQKSSGGPEAAAFVKVSMDGAPYLRKIDLRMYK"""
                     /note="color: #0080ff"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     primer_bind     complement(677..701)
                     /label=(1354) mAID seq 2
                     /note="color: black; sequence: ATCTTTAGGACAAGCACTCTTCTCC; 
                     added: 2021-03-06"
     CDS             885..1592
                     /codon_start=1
                     /label=mCherry2
                     /label=ORF frame 2
                     /note="color: #ff2132"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     gene            888..1592
                     /gene="mCherry (variant)"
                     /label=mCherry (variant)
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             1599..1655
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99ff
                     Cleavage site after base 1652"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1662..2060
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD (Aspergillus terreus cytidine deaminase)
                     /note="confers resistance to blasticidin"
                     /note="color: #ccffcc"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     1915..1937
                     /label=(1242) BSD seq1
                     /note="color: black; sequence: GCATCTTGAGCCCCTGCGGACGG; 
                     added: 2021-03-06"
     polyA_signal    2084..2539
                     /label=mouse PGK terminator
                     /note="color: #ffffff"
     protein_bind    complement(2563..2596)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     misc_feature    2563..2596
                     /label=LoxP-2
                     /note="color: #84a4c0; direction: RIGHT"
     promoter        complement(2645..2663)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2684..2700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2708..2724
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2732..2762)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2732 .. 2738 / #ffffff / -10
                        2: 2739 .. 2756 / #ffffff
                        3: 2757 .. 2762 / #ffffff / -35"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(3086..3674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(3845..4705)
                     /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: 3845 .. 4636 / #ccffcc
                        2: 4637 .. 4705 / #ccffcc / signal sequence
                     Cleavage site after base 4636"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4706..4810)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      join(4837..4839,1..453)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
       61 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      121 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      181 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      241 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      301 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      361 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      421 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
      481 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      541 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      601 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattggagc tccccgggat
      661 ccggtgcagg cgccaaggag aagagtgctt gtcctaaaga tccagccaaa cctccggcca
      721 aggcacaagt tgtgggatgg ccaccggtga gatcataccg gaagaacgtg atggtttcct
      781 gccaaaaatc aagcggtggc ccggaggcgg cggcgttcgt gaaggtatca atggacggag
      841 caccgtactt gaggaaaatc gatttgagga tgtataaagc tagcatggtg agcaagggcg
      901 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      961 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1021 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1081 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1141 actacttgaa gctgtccttc cccgagggct tcaattggga gcgcgtgatg aacttcgagg
     1201 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     1261 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag tgtcgtacca
     1321 tgggctggga ggcctccact gagcggatgt accccgagga cggcgccctg aagggcgaga
     1381 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     1441 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcgacatc aagttggaca
     1501 tcctttccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     1561 actccaccgg cggcatggac gagctgtaca agacgcgtgc tactaacttc agcctgctga
     1621 agcaggctgg agacgtggag gagaaccctg gacctctcga gatggccaag cctttgtctc
     1681 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     1741 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     1801 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     1861 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     1921 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     1981 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     2041 gttatgtgtg ggagggctaa tcgatgatat cagatccccg ggatgcagaa attgatgatc
     2101 tattaaacaa taaagatgtc cactaaaatg gaagtttttc ctgtcatact ttgttaagaa
     2161 gggtgagaac agagtaccta cattttgaat ggaaggattg gagctacggg ggtgggggtg
     2221 gggtgggatt agataaatgc ctgctcttta ctgaaggctc tttactattg ctttatgata
     2281 atgtttcata gttggatatc ataatttaaa caagcaaaac caaattaagg gccagctcat
     2341 tcctcccact catgatctat agatctatag atctctcgtg ggatcattgt ttttctcttg
     2401 attcccactt tgtggttcta agtactgtgg tttccaaatg tgtcagtttc atagcctgaa
     2461 gaacgagatc agcagcctct gttccacata cacttcattc tcagtattgt tttgccaagt
     2521 tctaattcca tcagaagctg gtcgagatcc ggaaccctta atataacttc gtataatgta
     2581 tgctatacga agttattagg tccctcgaag aggttcacta ggatccggta cccagctttt
     2641 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     2701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4801 gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
