LOCUS       Exported                5141 bp ds-DNA     circular SYN 24-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5141)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 24, 2022 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
COMMENT     LOCUS Exported 4731 bp ds-DNA circular SYN 21-MAY-2019 DEFINITION 
            Localization: C1 Cloning Vector, Excitation: 384, Emission: 450. 
            ACCESSION . VERSION . KEYWORDS mAzurite-C1 SOURCE synthetic DNA 
            construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 
            4731) AUTHORS Mena MA, Treynor TP, Mayo SL, Daugherty PS TITLE Blue 
            fluorescent proteins with enhanced brightness and photostability 
            from a structurally targeted library. JOURNAL Nat Biotechnol. 2006 
            Dec . 24(12):1569-71. PUBMED 17115054 REFERENCE 2 (bases 1 to 4731) 
            AUTHORS . TITLE Direct Submission JOURNAL Exported May 21, 2019 from
            SnapGene Server 1.1.58 http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5141
                     /organism="synthetic DNA construct"
                     /label=synthetic DNA construct
     enhancer        133..436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        437..640
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     591..611
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    677..681
                     /label=Kozak
                     /note="Kozak"
     CDS             682..1398
                     /codon_start=1
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1427..1576
                     /label=biRhoBAST
                     /note="RBRD (F30 based)"
     misc_feature    1583..1732
                     /label=biRhoBAST
                     /note="RBRD (tRNA based)"
     misc_feature    1831..1877
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2001..2122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2038..2057)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2092..2111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2129..2584)
                     /direction=LEFT
                     /label=LEFT
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2266..2287)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2478..2497
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2611..2715
                     /label=bla
     promoter        2717..3074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2740..2760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2925..3060
                     /direction=RIGHT
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2987..3006
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3109..3903
                     /codon_start=1
                     /label=aph(3')-II (or nptII)
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3163..3182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3773..3792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4091..4110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4135..4182
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4511..5099
                     /direction=RIGHT
                     /label=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5000..5019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      661 ccgctagcga attccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      721 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      781 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      841 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      901 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      961 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1021 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1081 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1141 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1201 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1261 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     1321 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1381 atggacgagc tgtacaagtc cggactcaga tctcgacggg tctagaggta tgtgggagga
     1441 acctccgcga aagcggtgaa ggagaggcgc aaggttaacc gcctcaggtt ccaggaggat
     1501 tcgtcctcca aaggcctccg cgaaagcggt gaaggagcgc cacaaggtta actggcgcag
     1561 gccttaccca cataccgcta gaggagcccg gaaggaacct ccgcgaaagc ggtgaaggag
     1621 aggcgcaagg ttaaccgcct caggttccag cggagttcgc tccgcaaagg cctccgcgaa
     1681 agcggtgaag gagcgccaca aggttaactg gcgcaggcct tatccgggct ccgctagagc
     1741 ggacttcggt ccgcttttta ctaggacctg caggcatgca agcttgacgt cggttaccga
     1801 tatccatatg gcggccgcat cgatctcgag ctcaagcttc gaattctgca gtcgacggta
     1861 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     1921 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1981 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2041 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2101 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     2161 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2221 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2281 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2341 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2401 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2461 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2521 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2581 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2641 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2701 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2821 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2881 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2941 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3001 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3061 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     3661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4081 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4141 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4201 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4261 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4321 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     4381 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4621 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5101 acgccagcaa cgcggccttt ttacggttcc tggccttttg c
//
