LOCUS       sequence-204494-        7291 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 7291)
  AUTHORS   Taylor MJ, Perrais D, Merrifield CJ
  TITLE     A high precision survey of the molecular dynamics of Mammalian 
            clathrin-mediated endocytosis.
  JOURNAL   PLoS Biol. 2011 Mar . 9(3):e1000604.
  PUBMED    21445324
REFERENCE   2  (bases 1 to 7291)
  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: "PLoS Biol. 
            2011 Mar . 9(3):e1000604."
COMMENT     Sequence Label: AP2u2-mCherry
FEATURES             Location/Qualifiers
     source          1..7291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      924..933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1416..2123
                     /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(1561..1579)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1829..1848
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2833..3062
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3045..3064
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3135..3709,3710..3712,3713..3721)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3302..3319)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3529..3548
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3744..4547
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3807..3826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4417..4436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4677..4798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4714..4733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4900..4916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4900..4916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4913..4935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4924..4940
                     /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(4948..4954,4955..4972,4973..4978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4993..5014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5131..5148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5302..5890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5382..5401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6061..6852,6853..6921))
                     /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     6684..6703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6922..7026)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7101..7120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgat atctgcggcc tagctagcac catgatcgga ggcttattca
      721 tctataatca caagggggag gtgcttatct cccgagtcta cagagatgac atcgggagga
      781 atgcagtgga tgcctttcgc gtcaatgtca ttcacgcacg gcagcaggtg cgcagccctg
      841 tcacaaacat cgctcgcacc agcttcttcc atgttaagcg gtccaacatc tggctggcag
      901 cagtcaccaa gcagaatgtc aatgccgcca tggtcttcga attcctctac aagatgtgtg
      961 atgtaatggc tgcttacttt ggcaagatca gcgaggagaa catcaagaac aatttcgtgc
     1021 tcatatacga gctgctggac gagattctgg actttggcta cccacagaac tcagagacag
     1081 gtgcactgaa aaccttcatc acccaacagg gtatcaagag tcagcatcag acaaaagaag
     1141 agcagtccca gatcaccagc caggtgaccg ggcaaattgg ctggaggcgg gaaggcatca
     1201 agtatcgtcg gaatgaactc ttcctagatg ttctggagag tgtgaacctg cttatgtccc
     1261 cacaggggca ggtgctgagt gcccatgtgt cgggccgggt ggtgatgaag agttacctga
     1321 gcggcatgcc tgagtgcaag tttggaatga atgacaagat tgtcatagaa aagcagggca
     1381 aaggcacagc cgatgaaaca agcaagagta gatctatggt gagcaagggc gaggaggata
     1441 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1501 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1561 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1621 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1681 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1741 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1801 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1861 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     1921 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     1981 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2041 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2101 gcggcatgga cgagctgtac aagcatatgg gtaagcagtc gatcgccatt gatgactgca
     2161 cctttcacca gtgtgtgcga ctcagcaagt ttgactctga gcggagcatc agctttatcc
     2221 ctcctgatgg agagtttgaa ctcatgagat accgtactac caaggacatc atcctgcctt
     2281 tccgggtgat cccattggtg cgggaggtgg gacgcaccaa actggaggtc aaggtggtca
     2341 tcaagtccaa cttcaagccc tcacttctgg cccagaagat tgaggtgagg atcccaaccc
     2401 cattgaacac aagcggggtg caagtgatct gcatgaaggg gaaggccaag tacaaggcca
     2461 gcgagaacgc cattgtatgg aagatcaagc gcatggcagg catgaaggaa tcacagatca
     2521 gcgcagagat tgagctcctg cccaccaatg ataagaagaa atgggctcgg ccccccattt
     2581 ccatgaactt tgaggtgcca tttgcgcctt ctggcctcaa ggtgcgctac ttgaaggtat
     2641 ttgagccgaa gctgaactac agcgaccacg atgtcatcaa atgggtgcgc tacattggcc
     2701 gaagtggcat ttacgaaacc cgctgctaga ccggtcgtac gtctccggat tcgaattcgg
     2761 atccgcggcc gcatagataa ctgatccagt gtgctggaat taattcgctg tctgcgaggg
     2821 ccagctgttg gggtgagtac tccctctcaa aagcgggcat gacttctgcg ctaagattgt
     2881 cagtttccaa aaacgaggag gatttgatat tcacctggcc cgcggtgatg cctttgaggg
     2941 tggccgcgtc catctggtca gaaaagacaa tctttttgtt gtcaagcttg aggtgtggca
     3001 ggcttgagat ctggccatac acttgagtga caatgacatc cactttgcct ttctctccac
     3061 aggtgtccac tcccaggtcc aactgcaggt cgagcatgca tctagggcgg ccaattccgc
     3121 ccctctcccc cccccccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     3181 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     3241 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     3301 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     3361 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     3421 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     3481 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     3541 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     3601 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     3661 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac
     3721 ccgggataat tcctgcagcc aatatgggat cggccattga acaagatgga ttgcacgcag
     3781 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3841 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3901 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3961 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4021 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4081 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4141 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4201 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4261 tgttcgccag gctcaaggcg cgcatgcccg acggcgatga tctcgtcgtg acccatggcg
     4321 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4381 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4441 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4501 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaggg gatcaattct
     4561 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4621 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4681 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4741 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     4801 gcgtcgagtg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4861 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6061 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6121 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6181 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6241 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6301 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6361 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6421 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6481 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6541 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6601 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6661 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6721 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6781 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6841 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6901 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6961 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7021 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7081 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7141 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7201 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     7261 cttcgcgatg tacgggccag atatacgcgt t
//
