LOCUS       Exported                9419 bp ds-DNA     circular SYN 23-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9419)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9419
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    1..225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(235..942)
                     /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"
     CDS             complement(943..996)
                     /codon_start=1
                     /label=T2A
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(997..1593)
                     /codon_start=1
                     /label=PuroR
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        1602..1931
                     /label=SV40 promoter
     enhancer        1939..2318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2014..3609
                     /label=CAG promoter
     promoter        2321..2596
                     /label=chicken beta-actin promoter
     promoter        3670..3688
                     /label=T7 promoter
     misc_feature    3695..3794
                     /label=LHA
     5'UTR           3795..3969
     polyA_signal    3976..4024
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    complement(4188..4373)
                     /label=GFP
     misc_feature    4374..5273
                     /label=intron
     misc_feature    complement(5274..5807)
                     /label=GFP-split
     promoter        complement(5880..6083)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        6084..6387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     3'UTR           6394..6718
     misc_feature    6719..6818
                     /label=RHA
     polyA_signal    6867..6988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7031..7047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7055..7071
                     /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(7079..7109)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(7433..8021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8192..9052)
                     /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"
     promoter        complement(9053..9157)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
       61 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
      121 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
      181 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagggatc cttacttgta
      241 cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat
      301 ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt aggcgccggg
      361 cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc
      421 gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct cggggtacat
      481 ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg
      541 gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc
      601 ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc
      661 ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt
      721 ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg ggccaccctt
      781 ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc
      841 gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc
      901 cttgatgatg gccatgttat cctcctcgcc cttgctcacc attgggccgg gattctcctc
      961 cacgtcaccg catgttagaa gacttcctct gccctcggca ccgggcttgc gggtcatgca
     1021 ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc
     1081 gtagaagggg aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc
     1141 ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg
     1201 cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag
     1261 gccttccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc
     1321 gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc
     1381 ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag
     1441 ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc
     1501 ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc cgggggacgt cgtcgcgggt
     1561 ggcgaggcgc accgtgggct tgtactcggt catggtcgcg ttttgcaaaa gcctaggcct
     1621 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     1681 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     1741 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     1801 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     1861 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     1921 aactgacaca cacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1981 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2041 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2101 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggactattta cggtaaactg
     2161 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2221 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2281 ggcagtacat ctacgtatta gtcatcgcta ttaccatggg tcgaggtgag ccccacgttc
     2341 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     2401 taattatttt gtgcagcgat gggggcgggg gggggggggg cgcgcgccag gcggggcggg
     2461 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     2521 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     2581 aagcgcgcgg cgggcgggag tcgctgcgtt gccttcgccc cgtgccccgc tccgcgccgc
     2641 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     2701 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggctcg tttcttttct
     2761 gtggctgcgt gaaagcctta aagggctccg ggagggccct ttgtgcgggg gggagcggct
     2821 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggcccg cgctgcccgg
     2881 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcgtgt gcgcgagggg
     2941 agcgcggccg ggggcggtgc cccgcggtgc gggggggctg cgaggggaac aaaggctgcg
     3001 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac
     3061 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     3121 cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
     3181 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc
     3241 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     3301 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     3361 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     3421 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
     3481 cggggctgtc cgcaggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     3541 ttctggcgtg tgaccggcgg ctctagcgcc tctgctaacc atgttcatgc cttcttcttt
     3601 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagct
     3661 agtgaattct aatacgactc actataggac cggtgttgac gcgatgtgat ttctgcccag
     3721 tgctctgaat gtcaaagtga agaaattcaa tgaagcgcgg gtaaacggcg ggagtaacta
     3781 tgactctctt aagggtctag ttacaactgg gcatcgctgc agagatcgca cctcctcgtg
     3841 gtcccgctgg tagcccttcg aagggtgact aagtcgatct ctgccccagg tacggagccg
     3901 ttgggactca ccagtccaac gtaactcctg cctaaattcg gtgaaacaaa ttcctcggta
     3961 aaaagccccg gtaccgaaca aacgacccaa cacccgtgcg ttttattctg tctttttatt
     4021 gccgatcccc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4081 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4141 caatatcacg ggtagccaac gctatgtcct gatagcggtc ggccgcttta cttgtacagc
     4201 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
     4261 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc
     4321 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc caggtgagtc
     4381 caggagatgt ttcagcactg ttgcctttag tctcgaggca acttagacaa ctgagtattg
     4441 atctgagcac agcagggtgt gagctgtttg aagatactgg ggttgggggt gaagaaactg
     4501 cagaggacta actgggctga gacccagtgg caatgtttta gggcctaagg aatgcctctg
     4561 aaaatctaga tggacaactt tgactttgag aaaagagagg tggaaatgag gaaaatgact
     4621 tttctttatt agatttcggt agaaagaact ttcatctttc ccctattttt gttattcgtt
     4681 ttaaaacatc tatctggagg caggacaagt atggtcatta aaaagatgca ggcagaaggc
     4741 atatattggc tcagtcaaag tggggaactt tggtggccaa acatacattg ctaaggctat
     4801 tcctatatca gctggacaca tataaaatgc tgctaatgct tcattacaaa cttatatcct
     4861 ttaattccag atgggggcaa agtatgtcca ggggtgagga acaattgaaa catttgggct
     4921 ggagtagatt ttgaaagtca gctctgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgcg
     4981 cgcacgtgtg tttgtgtgtg tgtgagagcg tgtgtttctt ttaacgtttt cagcctacag
     5041 catacagggt tcatggtggc aagaagataa caagatttaa attatggcca gtgactagtg
     5101 ctgcaagaag aacaactacc tgcatttaat gggaaagcaa aatctcaggc tttgagggaa
     5161 gttaacatag gcttgattct gggtggaagc tgggtgtgta gttatctgga ggccaggctg
     5221 gagctctcag ctcactatgg gttcatcttt attgtctcct ttcatctcaa cagctgcacg
     5281 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
     5341 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
     5401 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     5461 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     5521 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     5581 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     5641 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     5701 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     5761 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaattcg
     5821 aagcttgagc tcgagatctg agtccggtag cgctagcgga tctgacggtt cactaaacca
     5881 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     5941 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     6001 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     6061 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     6121 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     6181 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     6241 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     6301 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     6361 gggccattta ccgtaagtta tgtaacgccc gggttcaggt tatttagatg cttagttttt
     6421 gtacctttct tgttttgttt aggattttga tagtgttagt atttttatat ttttgtacga
     6481 ttgcataatg ttctttttta tacagttctg ttttaataaa atagacgata gctagagacg
     6541 ttagggcagc cacaagccag ttaggtagcg gatagtaggt aggaacagac ttttactatt
     6601 tcataacgcg tcaattacca cctgatttgg accaattcac gggatttgtc caaggtggac
     6661 gggccacctt tacttaaccc ggaaaaggaa catatataat ttatgtgtgt tcgataaata
     6721 gccaaatgcc tcgtcatcta attagtgacg cgcatgaatg gatgaacgag attcccactg
     6781 tccctaccta ctatccagcg aaaccacagc caagggaagc tagcctgcag cgcggggatc
     6841 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6961 gtttgtccaa actcatcaat gtatcttatt aattaacgtc gacctctagc tagagcttgg
     7021 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7081 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     7141 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7861 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9181 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     9241 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     9301 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     9361 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatat
//
