LOCUS       Exported                7666 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEF6.mCherry-TSG101
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  AUTHORS   Nabhan JF, Hu R, Oh RS, Cohen SN, Lu Q
  TITLE     Formation and release of arrestin domain-containing protein 
            1-mediated microvesicles (ARMMs) at plasma membrane by recruitment 
            of TSG101 protein.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4146-51. Epub 2012 Feb
            6.
  PUBMED    22315426
REFERENCE   2  (bases 1 to 7666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7666
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        123..1301
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          354..1292
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1249..1269
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1318..1337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1318..1336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1381..2088
                     /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"
     primer_bind     complement(1526..1544)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1794..1813
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3320..3349
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3365..3382
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3405..3422)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3411..3635
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3681..4109
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3768..3787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3978..3999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4118..4138)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4123..4452
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4303..4438
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4365..4384
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4500..4547
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4566..4964
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5122..5243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5159..5178)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5213..5232
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5292..5308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5292..5308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5305..5327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5316..5332
                     /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(5340..5370)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5385..5406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5523..5540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5683..6271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5763..5782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6442..7302)
                     /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"
     primer_bind     7065..7084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7303..7407)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7482..7501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt
       61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa
     1321 tacgactcac tatagggaga cccaagctgg ctaggtaagc ttggtaccga gctcggatcc
     1381 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1441 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1501 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1561 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1621 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     1681 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1741 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1801 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1861 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1921 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1981 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     2041 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagat ggcggtgtcg
     2101 gagagccagc tcaagaaaat ggtgtccaag tacaaataca gagacctaac tgtacgtgaa
     2161 actgtcaatg ttattactct atacaaagat ctcaaacctg ttttggattc atatgttttt
     2221 aacgatggca gttccaggga actaatgaac ctcactggaa caatccctgt gccttataga
     2281 ggtaatacat acaatattcc aatatgccta tggctactgg acacataccc acataatccc
     2341 cctatctgtt ttgttaagcc tactagttca atgactatta aaacaggaaa gcatgttgat
     2401 gcaaatggga agatatatct tccttatcta catgaatgga aacacccaca gtcagacttg
     2461 ttggggctta ttcaggtcat gattgtggta tttggagatg aacctccagt cttctctcgt
     2521 cctatttcgg catcctatcc gccataccag gcaacggggc caccaaatac ttcctacatg
     2581 ccaggcatgc caggtggaat ctctccatac ccatccggat accctcccaa tcccagtggt
     2641 tacccaggct gtccttaccc acctggtggt ccatatcctg ccacaacaag ttctcagtac
     2701 ccttctcagc ctcctgtgac cactgttggt cccagtaggg atggcacaat cagcgaggac
     2761 accatccgag cctctctcat ctctgcggtc agtgacaaac tgagatggcg gatgaaggag
     2821 gaaatggatc gtgcccaggc agagctcaat gccttgaaac gaacagaaga agacctgaaa
     2881 aagggtcacc agaaactgga agagatggtt acccgtttag atcaagaagt agccgaggtt
     2941 gataaaaaca tagaactttt gaaaaagaag gatgaagaac tcagttctgc tctggaaaaa
     3001 atggaaaatc agtctgaaaa caatgatatc gatgaagtta tcattcccac agctccctta
     3061 tacaaacaga tcctgaatct gtatgcagaa gaaaacgcta ttgaagacac tatcttttac
     3121 ttgggagaag ccttgagaag gggcgtgata gacctggatg tcttcctgaa gcatgtacgt
     3181 cttctgtccc gtaaacagtt ccagctgagg gcactaatgc aaaaagcaag aaagactgcc
     3241 ggtctcagtg acctctactg agaattctgc agatatccag cacagtggcg gccgctcgag
     3301 tctagagggc ccgcggttcg aacaaaaact catctcagaa gaggatctga atatgcatac
     3361 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3421 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3481 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3541 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3601 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3661 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3721 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3781 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3841 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3901 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3961 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4021 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4081 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4261 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4321 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4381 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4441 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     4501 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     4561 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     4621 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     4681 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     4741 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     4801 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     4861 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     4921 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag
     4981 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5041 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5101 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5161 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5221 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5281 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5341 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5401 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5461 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7441 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7501 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7561 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7621 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatata
//
