LOCUS       Exported                7570 bp ds-DNA     circular SYN 16-AUG-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7570)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Aug 16, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7570
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      46..1388
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
                     /note="color: #ffff00"
     protein_bind    378..425
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /note="color: #31849b; direction: RIGHT"
     promoter        1415..1519
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1520..2380
                     /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: 1520 .. 1588 / #ccffcc / signal sequence
                        2: 1589 .. 2380 / #ccffcc
                     Cleavage site after base 1588"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2551..3139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        3463..3493
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 3463 .. 3468 / #ffffff / -35
                        2: 3469 .. 3486 / #ffffff
                        3: 3487 .. 3493 / #ffffff / -10"
     protein_bind    3501..3517
                     /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"
     primer_bind     3506..3528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     primer_bind     3525..3541
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     3525..3541
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     3560..3580
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: black; sequence: GCAATTAACCCTCACTAAAGG"
     promoter        3562..3580
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    3598..4261
                     /label=GPD prom
                     /note="color: #00ffff"
     primer_bind     4141..4160
                     /label=seq-F
                     /note="color: black; sequence: gacggtaggtattgattgta; added:
                     2020-04-21"
     primer_bind     4255..4271
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="This forward directional feature has 2 segments:
                        1: 4255 .. 4267 / #a020f0
                        2: 4268 .. 4271 / #a020f0"
     primer_bind     4255..4271
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
                     /note="color: black; sequence: TCTAGAACTAGTGGATC"
     protein_bind    4386..4407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        4422..4452
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 4422 .. 4427 / #ffffff / -35
                        2: 4428 .. 4445 / #ffffff
                        3: 4446 .. 4452 / #ffffff / -10"
     protein_bind    4460..4476
                     /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"
     RBS             4494..4505
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /note="color: #a6acb3"
     CDS             4512..5189
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="color: #ff0000"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      5259..5506
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
     primer_bind     complement(5334..5351)
                     /label=seq-R
                     /note="color: black; sequence: CCTAGACTTCAGGTTGTC; added: 
                     2020-04-21"
     primer_bind     complement(5508..5516)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(5541..5560)
                     /label=T7
                     /note="T7 promoter, forward primer"
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        complement(5542..5560)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(5570..5587)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(5570..5586)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(5579..5601)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     rep_origin      5731..6186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(6446..7105)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /note="color: #ff7f50"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(7106..7292)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 cctattttta taggttaatg tcatgataat aatggtttct tagatgatcc aatatcaaag
       61 gaaatgatag cattgaagga tgagactaat ccaattgagg agtggcagca tatagaacag
      121 ctaaagggta gtgctgaagg aagcatacga taccccgcat ggaatgggat aatatcacag
      181 gaggtactag actacctttc atcctacata aatagacgca tataagtacg catttaagca
      241 taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata
      301 ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc
      361 gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctaga aagtatagga
      421 acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa
      481 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct
      541 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac
      601 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag
      661 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa
      721 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc
      781 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg
      841 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc
      901 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa
      961 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca
     1021 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga
     1081 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg
     1141 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
     1201 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
     1261 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
     1321 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
     1381 gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1441 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1501 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1561 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1621 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1681 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1741 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1801 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1861 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1921 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1981 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2041 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2101 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2161 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2221 agccggtgag cgtggatctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2281 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2341 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2401 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2461 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2521 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2701 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2821 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2881 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2941 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3001 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3061 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3121 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3181 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3241 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3301 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3361 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3421 cgcaattaat gtgagttacc tcactcatta ggcaccccag gctttacact ttatgcttcc
     3481 ggctcctatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     3541 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctctc
     3601 gagtttatca ttatcaatac tgccatttca aagaatacgt aaataattaa tagtagtgat
     3661 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
     3721 caaaataggg ggcgggttac acagaatata taacactgat ggtgcttggg tgaacaggtt
     3781 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     3841 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     3901 tcttagcgca actacagaga acagggcaca aacaggcaaa aaacgggcac aacctcaatg
     3961 gagtgatgca acctgcctgg agtaaatgat gacacaaggc aattgaccca cgcatgtatc
     4021 tatctcattt tcttacacct tctattacct tctgctctct ctgatttgga aaaagctgaa
     4081 aaaaaaggtt taaaccagtt ccctgaaatt attcccctac ttgactaata agtatataaa
     4141 gacggtaggt attgattgta attctgtaaa tctatttctt aaacttctta aattctactt
     4201 ttatagttag tctttttttt agttttaaaa caccaagaac ttagtttcga cggatctaga
     4261 actagtggat ccccaaaatg agacccaata cgcaaaccgc ctctccccgc gcgttggccg
     4321 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4381 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     4441 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacatacta gagaaagagg
     4501 agaaatacta gatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc
     4561 gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc
     4621 cgtacgaagg tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg
     4681 cttgggacat cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg
     4741 ctgacatccc ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta
     4801 tgaacttcga agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg
     4861 agttcatcta caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc
     4921 agaaaaaaac catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc
     4981 tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag
     5041 ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca
     5101 tcaaactgga catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg
     5161 ctgaaggtcg tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac
     5221 tagagccagg cggtctctac tgggggctgc aggaattctc atgtaattag ttatgtcacg
     5281 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     5341 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     5401 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     5461 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcgtcg acctcgaggg
     5521 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     5581 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5641 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5701 agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg
     5761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5941 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6061 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6121 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     6181 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga
     6241 tccgtcgagt tcaagagaaa aaaaaagaaa aagcaaaaag aaaaaaggaa agcgcgcctc
     6301 gttcagaatg acacgtatag aatgatgcat taccttgtca tcttcagtat catactgttc
     6361 gtatacatac ttactgacat tcataggtat acatatatac acatgtatat atatcgtatg
     6421 ctgcagcttt aaataatcgg tgtcactaca taagaacacc tttggtggag ggaacatcgt
     6481 tggtaccatt gggcgaggtg gcttctctta tggcaaccgc aagagccttg aacgcactct
     6541 cactacggtg atgatcattc ttgcctcgca gacaatcaac gtggagggta attctgctag
     6601 cctctgcaaa gctttcaaga aaatgcggga tcatctcgca agagagatct cctactttct
     6661 ccctttgcaa accaagttcg acaactgcgt acggcctgtt cgaaagatct accaccgctc
     6721 tggaaagtgc ctcatccaaa ggcgcaaatc ctgatccaaa cctttttact ccacgcgcca
     6781 gtagggcctc tttaaaagct tgaccgagag caatcccgca gtcttcagtg gtgtgatggt
     6841 cgtctatgtg taagtcacca atgcactcaa cgattagcga ccagccggaa tgcttggcca
     6901 gagcatgtat catatggtcc agaaacccta tacctgtgtg gacgttaatc acttgcgatt
     6961 gtgtggcctg ttctgctact gcttctgcct ctttttctgg gaagatcgag tgctctatcg
     7021 ctaggggacc accctttaaa gagatcgcaa tctgaatctt ggtttcattt gtaatacgct
     7081 ttactagggc tttctgctct gtcatctttg ccttcgttta tcttgcctgc tcatttttta
     7141 gtatattctt cgaagaaatc acattacttt atataatgta taattcatta tgtgataatg
     7201 ccaatcgcta agaaaaaaaa agagtcatcc gctaggggaa aaaaaaaaat gaaaatcatt
     7261 accgaggcat aaaaaaatat agagtgtact agaggaggcc aagagtaata gaaaaagaaa
     7321 attgcgggaa aggactgtgt tatgacttcc ctgactaatg ccgtgttcaa acgatacctg
     7381 gcagtgactc ctagcgctca ccaagctctt aaaacgggaa tttatggtgc actctcagta
     7441 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     7501 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg acaaagggcc
     7561 tcgtgatacg
//
