LOCUS       V022732                 8135 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022732
VERSION     V022732
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..2938
                     /label="GLB1(NM_000404)"
                     /note="GLB1(NM_000404)"
                     /gene="GLB1"
     CDS             2954..3655
                     /codon_start=1
                     /product="monomeric cyan-excitable red fluorescent protein
                     (Laviv et al., 2016)"
                     /transl_table=1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTA
                     RIKVVEGGPLPFAFDILATMFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKLRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KKLKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
                     /label="mCyRFP1"
     polyA_signal    3702..3926
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3972..4400
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4414..4743
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4810..5604
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5778..5899
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5948..5964)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5972..5988
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5996..6026)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6041..6062
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6350..6935)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7106..7966)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7967..8071)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg ccggggttcc tggttcgcat cctccctctg ttgctggttc tgctgcttct
      961 gggccctacg cgcggcttgc gcaatgccac ccagaggatg tttgaaattg actatagccg
     1021 ggactccttc ctcaaggatg gccagccatt tcgctacatc tcaggaagca ttcactactc
     1081 ccgtgtgccc cgcttctact ggaaggaccg gctgctgaag atgaagatgg ctgggctgaa
     1141 cgccatccag acgtatgtgc cctggaactt tcatgagccc tggccaggac agtaccagtt
     1201 ttctgaggac catgatgtgg aatattttct tcggctggct catgagctgg gactgctggt
     1261 tatcctgagg cccgggccct acatctgtgc agagtgggaa atgggaggat tacctgcttg
     1321 gctgctagag aaagagtcta ttcttctccg ctcctccgac ccagattacc tggcagctgt
     1381 ggacaagtgg ttgggagtcc ttctgcccaa gatgaagcct ctcctctatc agaatggagg
     1441 gccagttata acagtgcagg ttgaaaatga atatggcagc tactttgcct gtgattttga
     1501 ctacctgcgc ttcctgcaga agcgctttcg ccaccatctg ggggatgatg tggttctgtt
     1561 taccactgat ggagcacata aaacattcct gaaatgtggg gccctgcagg gcctctacac
     1621 cacggtggac tttggaacag gcagcaacat cacagatgct ttcctaagcc agaggaagtg
     1681 tgagcccaaa ggacccttga tcaattctga attctatact ggctggctag atcactgggg
     1741 ccaacctcac tccacaatca agaccgaagc agtggcttcc tccctctatg atatacttgc
     1801 ccgtggggcg agtgtgaact tgtacatgtt tataggtggg accaattttg cctattggaa
     1861 tggggccaac tcaccctatg cagcacagcc caccagctac gactatgatg ccccactgag
     1921 tgaggctggg gacctcactg agaagtattt tgctctgcga aacatcatcc agaagtttga
     1981 aaaagtacca gaaggtccta tccctccatc tacaccaaag tttgcatatg gaaaggtcac
     2041 tttggaaaag ttaaagacag tgggagcagc tctggacatt ctgtgtccct ctgggcccat
     2101 caaaagcctt tatcccttga catttatcca ggtgaaacag cattatgggt ttgtgctgta
     2161 ccggacaaca cttcctcaag attgcagcaa cccagcacct ctctcttcac ccctcaatgg
     2221 agtccacgat cgagcatatg ttgctgtgga tgggatcccc cagggagtcc ttgagcgaaa
     2281 caatgtgatc actctgaaca taacagggaa agctggagcc actctggacc ttctggtaga
     2341 gaacatggga cgtgtgaact atggtgcata tatcaacgat tttaagggtt tggtttctaa
     2401 cctgactctc agttccaata tcctcacgga ctggacgatc tttccactgg acactgagga
     2461 tgcagtgtgc agccacctgg ggggctgggg acaccgtgac agtggccacc atgatgaagc
     2521 ctgggcccac aactcatcca actacacgct cccggccttt tatatgggga acttctccat
     2581 tcccagtggg atcccagact tgccccagga cacctttatc cagtttcctg gatggaccaa
     2641 gggccaggtc tggattaatg gctttaacct tggccgctat tggccagccc ggggccctca
     2701 gttgaccttg tttgtgcccc agcacatcct gatgacctcg gccccaaaca ccatcaccgt
     2761 gctggaactg gagtgggcac cctgcagcag tgatgatcca gaactatgtg ctgtgacgtt
     2821 cgtggacagg ccagttattg gctcatctgt gacctacgat catccctcca aacctgttga
     2881 aaaaagactc atgcccccac ccccgcaaaa aaacaaagat tcatggctgg accatgtagg
     2941 cggaggtgga agtatggtga gcaagggcga ggagctgatc aaggagaaca tgagaagcaa
     3001 gctgtacctg gaaggcagcg tgaacggcca ccagttcaag tgcacccacg aaggggaggg
     3061 caagccctac gagggcaagc agaccgcgag gatcaaggtg gtggagggag gccccctgcc
     3121 gttcgcattc gacatcctgg ccaccatgtt tatgtacggg agcaaggtgt tcatcaagta
     3181 ccccgccgac ctccccgatt attttaagca gtccttccct gagggcttca catgggagag
     3241 agtcatggtg ttcgaagacg ggggcgtgct gaccgccacc caggacacca gcctccagga
     3301 cggcgagctc atctacaacg tcaagctcag aggggtgaac ttcccagcca acggccccgt
     3361 gatgcagaag aaaacactgg gctgggagcc cagcaccgag accatgtacc ccgctgacgg
     3421 cggcctggaa ggcagatgcg acaagaagct gaagctcgtg ggcgggggcc acctgcacgt
     3481 caacttcaag accacataca agtccaagaa acccgtgaag atgcccggcg tccactacgt
     3541 ggaccgcaga ctggaaagaa tcaaggaggc cgacaacgag acctacgtcg agcagtacga
     3601 gcacgctgtg gccagatact ccaacctggg cggaggcatg gacgagctgt acaagtagct
     3661 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3721 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3781 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3841 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3901 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3961 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4021 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4081 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4141 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4201 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4261 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4321 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4381 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4501 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4561 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4621 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4681 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4741 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4801 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4861 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4921 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4981 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5041 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5101 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5161 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5221 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5281 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5341 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5401 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5461 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5521 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5581 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5641 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5701 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5761 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5821 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5881 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5941 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6001 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6061 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6121 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6181 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6241 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6301 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6361 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6421 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6481 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6541 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6601 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6661 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6721 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6781 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6841 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6901 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6961 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7021 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7081 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7141 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7201 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7261 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7321 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7381 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7441 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7501 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7561 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7621 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7681 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7741 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7801 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7861 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7921 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7981 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8041 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8101 tcgacggatc gggagatctc ccgatcccct atggt
//
