LOCUS       sequence-48383-e        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2012-03-13"; volume: "109(11)"; pages: 
            "4146-51. Epub 2012 Feb 6"
COMMENT     Sequence Label: pEF6.mCherry-TSG101
FEATURES             Location/Qualifiers
     source          1..7666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             58..87
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             103..120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(143..160)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    149..373
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      419..847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(506..525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     716..737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(856..876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        861..1190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1041..1176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1103..1122
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1238..1285
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1304..1702
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1860..1981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1897..1916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1951..1970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2030..2046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2043..2065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2054..2070
                     /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(join(2078..2084,2085..2102,2103..2108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2123..2144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2261..2278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2421..3009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2501..2520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3180..3971,3972..4040))
                     /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     3803..3822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4041..4145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4220..4239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4527..5705
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4758..5696
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5653..5673
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     5722..5741
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5722..5740
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5785..6492
                     /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(5930..5948)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6198..6217
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc gcggttcgaa
       61 caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat
      121 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
      181 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
      241 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
      301 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
      361 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
      421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      481 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      541 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      601 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      721 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      781 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      841 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
      901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
      961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1021 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1081 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1141 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1201 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     1261 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc
     1321 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     1381 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     1441 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     1501 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     1561 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     1621 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     1681 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac
     1741 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     1801 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     1861 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1921 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1981 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     2041 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag gaaccgtaaa
     2401 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2461 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2521 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2581 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2641 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2701 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2761 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2821 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2881 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2941 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3001 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3061 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3121 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3301 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3481 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3661 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4021 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4141 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4201 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4261 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4321 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4381 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagg cttttgcaaa
     4441 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     4501 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     4561 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     4621 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4681 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4741 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     4801 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     4861 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     4921 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     4981 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     5041 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     5101 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     5161 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     5221 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     5281 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     5341 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     5401 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     5461 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     5521 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     5581 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     5641 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     5701 cgtgaggaat tagcttggta ctaatacgac tcactatagg gagacccaag ctggctaggt
     5761 aagcttggta ccgagctcgg atccatggtg agcaagggcg aggaggataa catggccatc
     5821 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     5881 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     5941 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     6001 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     6061 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     6121 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     6181 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     6241 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     6301 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     6361 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     6421 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     6481 gagctgtaca agatggcggt gtcggagagc cagctcaaga aaatggtgtc caagtacaaa
     6541 tacagagacc taactgtacg tgaaactgtc aatgttatta ctctatacaa agatctcaaa
     6601 cctgttttgg attcatatgt ttttaacgat ggcagttcca gggaactaat gaacctcact
     6661 ggaacaatcc ctgtgcctta tagaggtaat acatacaata ttccaatatg cctatggcta
     6721 ctggacacat acccatataa tccccctatc tgttttgtta agcctactag ttcaatgact
     6781 attaaaacag gaaagcatgt tgatgcaaat gggaagatat atcttcctta tctacatgaa
     6841 tggaaacacc cacagtcaga cttgttgggg cttattcagg tcatgattgt ggtatttgga
     6901 gatgaacctc cagtcttctc tcgtcctatt tcggcatcct atccgccata ccaggcaacg
     6961 gggccaccaa atacttccta catgccaggc atgccaggtg gaatctctcc atacccatcc
     7021 ggataccctc ccaatcccag tggttaccca ggctgtcctt acccacctgg tggtccatat
     7081 cctgccacaa caagttctca gtacccttct cagcctcctg tgaccactgt tggtcccagt
     7141 agggatggca caatcagcga ggacaccatc cgagcctctc tcatctctgc ggtcagtgac
     7201 aaactgagat ggcggatgaa ggaggaaatg gatcgtgccc aggcagagct caatgccttg
     7261 aaacgaacag aagaagacct gaaaaagggt caccagaaac tggaagagat ggttacccgt
     7321 ttagatcaag aagtagccga ggttgataaa aacatagaac ttttgaaaaa gaaggatgaa
     7381 gaactcagtt ctgctctgga aaaaatggaa aatcagtctg aaaacaatga tatcgatgaa
     7441 gttatcattc ccacagctcc cttatacaaa cagatcctga atctgtatgc agaagaaaac
     7501 gctattgaag acactatctt ttacttggga gaagccttga gaaggggcgt gatagacctg
     7561 gatgtcttcc tgaagcatgt acgtcttctg tcccgtaaac agttccagct gagggcacta
     7621 atgcaaaaag caagaaagac tgccggtctc agtgacctct actgag
//
