LOCUS       sequence-148012-        6493 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 6493)
  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 6493)
  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: SNX9-pmCherryC1
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             607..1314
                     /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(752..770)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1020..1039
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3288..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3325..3344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3379..3398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3416..3871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3553..3574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3765..3784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3898..4002
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4004..4361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4004..4195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4027..4047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4212..4347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4396..5190
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4396..5190
                     /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(4450..4469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5060..5079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5378..5397)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5422..5469
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5798..6386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6287..6306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      661 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      781 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      841 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1021 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1081 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1141 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1201 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1261 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga
     1321 ctcagatcta ccatggccac caaggctcgg gtcatgtacg actttgctgc tgagcctgga
     1381 aacaatgagc tgaccgttac tgaaggagaa atcatcacag ttacaaaccc gaatgttggt
     1441 ggaggatggc tggaaggaaa gaacaacaaa ggagagcagg ggcttgttcc tactgactac
     1501 gtggaaattt taccaaacga tggaaaagat ccgttttctt gtgggaattc agtagccgac
     1561 caagctttcc tcgattccct gacagctagc acagctcaaa ccaactcatc ttctgccaac
     1621 agcaataacc aggttggtgg tggcaatgac ccctggacag cctggaatgc tcccaaacct
     1681 gggaactggg acagctccga tgcctggggc tccaggacag acggaactag cgctcagagg
     1741 aacagcagcg ccaacaactg ggacacgggc tttggccacc cacaggccta ccaaggacca
     1801 gcaactggtg atgatgacga atgggatgaa gactgggacg accccaagtc ctcctcacct
     1861 tacttcaagg attcagaacc agctgaagca gggggcatcc agcggggaaa cagtcgtgcg
     1921 ggtgcctcct ccatgaagct gccccttaac aaatttcctg gatttgcaaa acctggcatg
     1981 gagcagtact tgttggccaa gcagctagca aaacccaaag agaaaatcgc catcatcgtt
     2041 ggagattatg gccctatgtg ggtttatcct acctctacgt ttgactgtgt ggtagctgat
     2101 ccccggaaag gttccaaaat gtacggtctg aagagctaca ttgagtatca gctaacaccc
     2161 actaacacta accgatctgt aaaccacaga tataagcact ttgattggtt atatgagcgt
     2221 ctcctggtta agtttgggtc tgccattccg atcccttctc ttccagacaa gcaggtcaca
     2281 ggtcgctttg aagaggaatt tatcaaaatg cggatggagc ggctccaggc ctggatgacc
     2341 aggatgtgcc ggcacccggt ggtgtcagaa agtgaagttt tccagcagtt cctgaatttc
     2401 cgagatgaga aggaatggaa gactggaaag aggaaggctg agaaagacga actggtggga
     2461 gtgatgatat tttccaccat ggaaccagag gctcctgact tggatttgat agaaatagaa
     2521 cagaagtgtg acgctgtggg gaagttcacc aaggccatgg atgacggcgt gaaggagctg
     2581 ctgacggtgg ggcaggagca ctggaagcgc tgcacagggc ccttacccaa ggagtatcag
     2641 aagataggga aggctctgca gagcttagcg gcagtgttca gctccagcgg ttatcaagga
     2701 gagaccgacc ttaacgatgc aatcacagaa gctgggaaga cgtatgagga gatcgccagc
     2761 cttgtggcag agcagccaaa gaaggacctc cacttcctga tggagtgcaa ccatgagtac
     2821 aaaggctttc tcggctgctt cccggacatc attggtgccc acaagggagc aatagaaaaa
     2881 gtgaaagaaa gtgataagct ggttgcaact agtaaaatta ctccccaaga caagcagacc
     2941 atggtgaaga gagttggcac catgtcttac gctctgcaag ctgagatgaa tcactttcac
     3001 agtaaccgga tctatgatta caacagtgtc atccgcctct acctggagca gcaagtgcag
     3061 ttctatgaaa caatcgcaga aaagctgagg caggccctca gccgcttccc ggttatgggt
     3121 tcttcaggat catccgggtc gacggtaccg cgggcccggg atccaccgga tctagataac
     3181 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3241 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     3421 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3481 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3541 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3601 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3661 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3721 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3781 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3841 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3901 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3961 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4021 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4201 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4261 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4321 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4381 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4441 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4501 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4561 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4621 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4681 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4741 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4801 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4861 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4921 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4981 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5041 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5101 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5161 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5221 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5281 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5341 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5401 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5461 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5521 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5581 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5641 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5701 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5941 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6421 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6481 accgtattac cgc
//
