LOCUS       Exported                5761 bp ds-DNA     circular SYN 16-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5761)
  AUTHORS   Joe
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 4, 2020 from SnapGene 5.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5761
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     1..28
                     /label=Fragment 1.FOR
     CDS             11..70
                     /codon_start=1
                     /label=Membrane Trafficking signal
                     /note="Membrane trafficking signal"
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     complement(50..70)
                     /label=eViRho1_dC_r
     CDS             71..784
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(113..134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(121..132)
                     /label=Fragment 2.REV
     primer_bind     complement(374..393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     721..742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             785..808
                     /codon_start=1
                     /label=ER release signal
                     /translation="FCYENEV"
     primer_bind     806..838
                     /label=eViRho1_dC_f
                     /note="for gibson assembly to generate 1D4-tagged 
                     eViRho1_dC"
     polyA_signal    919..1040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1010..1029
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1047..1502)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1184..1205)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1396..1415
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1529..1633
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1635..1992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1658..1678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1843..1978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1905..1924
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(1928..1939)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2027..2821
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2081..2100)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2691..2710
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3009..3028)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3053..3100
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3429..4017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3918..3937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4192..4495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4496..4699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4650..4670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    4740..4796
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      4797..4806
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4803..4934
                     /codon_start=1
                     /label=N-Terminus eKR2
                     /translation="MDYGGALSAVGLFQTSYTLENNGSVICIPNNGQCFCLAWLKSNG"
     primer_bind     complement(4916..4944)
                     /label=Fragment 1.REV
     primer_bind     4925..4955
                     /label=Fragment 2.FOR
     CDS             join(4935..5761,1..10)
                     /codon_start=1
                     /label=viRho1_dC
                     /label=293	2623079_2164382
                     /translation="MVSDTRIRETLQVIDLLWSISLFAFFMYQKMCKNRCKFEVMYVSS
                     IGSITDLVKLAFTDYTPASITIKENDIVFNWSQYYGWLITCPVLLIHLSNLAGKDVFDV
                     RRMMKILVAYQILMLSGATASLSEKDSPVKWCFFILAISCLMILYRYAAHIFLESMKIM
                     PKNATSVLKQIAIVFYSSWSGFGFFWIIGPSGLALVSNEISKAGFAFCDILSKNIYSML
                     GWYLRWYVLRKHDKPDEFINQTETPLKKITILLVDNEHNFLHYFSTILYQLDCDIYV"
     primer_bind     complement(join(5750..5761,1..20))
                     /label=Fragment 2.REV
ORIGIN
        1 tatctacgtg aagagcagga tcaccagcga gggcgagtac atccccctgg accagatcga
       61 catcaacgtg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      121 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      181 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      241 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
      301 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      361 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      421 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      481 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      541 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      601 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
      661 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
      721 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
      781 caagttctgc tacgagaacg aggtgtgatc tagatcataa tcagccatac cacatttgta
      841 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
      901 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
      961 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1021 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1081 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1141 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1201 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1261 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1321 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     1381 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     1441 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     1501 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1561 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1621 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     1681 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1741 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1801 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     1861 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     1921 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     1981 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2041 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2101 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2161 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2221 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2281 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2341 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2401 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2461 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2521 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     2581 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2641 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2701 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2761 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     2821 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     2881 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     2941 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3001 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3061 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3241 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     3301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3541 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     4141 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4201 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4261 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4321 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     4381 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     4441 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     4501 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4561 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     4621 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     4681 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
     4741 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccgcca
     4801 ccatggacta tgggggagca ctgtccgccg tcgggctgtt tcagacttcc tacaccctgg
     4861 agaacaacgg gagcgtcatt tgcattccta acaatggcca gtgcttctgt ctggcctggc
     4921 tgaagagcaa cgggatggtc agcgacaccc gaatcaggga gacactgcag gtcatcgatc
     4981 tgctgtggtc catctcactg tttgctttct ttatgtatca gaagatgtgc aagaacaggt
     5041 gtaagtttga agtgatgtac gtgagctcca tcggctccat caccgacctg gtgaagctgg
     5101 ccttcaccga ttatacacct gcctctatca caatcaagga gaacgacatc gtgtttaatt
     5161 ggagccagta ctatggctgg ctgatcacct gcccagtgct gctgatccac ctgtctaatc
     5221 tggccggcaa ggacgtgttt gatgtgagga gaatgatgaa gatcctggtg gcctaccaga
     5281 tcctgatgct gtccggcgcc acagcctccc tgtctgagaa ggattctccc gtgaagtggt
     5341 gcttctttat cctggccatc agctgtctga tgatcctgta ccgctacgcc gcccacatct
     5401 tcctggagag catgaagatc atgcccaaga acgccacctc cgtgctgaag cagatcgcca
     5461 tcgtgttcta ctctagctgg agcggctttg gcttcttttg gatcatcgga ccttccggcc
     5521 tggccctggt gagcaacgag atctccaagg ccggcttcgc cttttgcgac atcctgtcca
     5581 agaatatcta ttctatgctg ggctggtatc tgaggtggta tgtgctgaga aagcacgaca
     5641 agccagatga gttcatcaac cagaccgaga cacccctgaa gaagatcacc atcctgctgg
     5701 tggataacga gcacaatttc ctgcactact tttccacaat cctgtatcag ctggactgcg
     5761 a
//
