LOCUS       sequence-246248-        6800 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 6800)
  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 6800)
  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: Amph1-pmCherryN1
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2690..2699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2696..3400
                     /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
                     EGRHSTGGMDELY"
     primer_bind     complement(2841..2859)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3109..3128
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3528..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3565..3584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3619..3638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3656..4111)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3793..3814)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4005..4024
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4138..4242
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4244..4601
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4244..4435
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4267..4287)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4452..4587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4636..5430
                     /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             4636..5430
                     /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(4690..4709)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5300..5319
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5618..5637)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5662..5709
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6038..6626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6527..6546
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcaccatg gccgacatca agacgggcat
      601 cttcgccaaa aacgtgcaaa agcgcctcaa ccgcgcgcag gaaaaggtcc tccagaagct
      661 ggggaaggct gacgagacaa aagatgaaca gtttgaagaa tatgtccaga acttcaaacg
      721 gcaggaggca gagggtacca gacttcagcg agaactccgg ggatatttag cagcaatcaa
      781 aggtatgcag gaggcctcaa tgaagctcac tgaatctcta catgaagtct acgagccaga
      841 ctggtatgga cgggaagatg taaagatggt tggtgagaag tgtgatgtgc tgtgggaaga
      901 cttccatcaa aagctagtgg atggctcctt gctgacactg gacacctacc tgggccagtt
      961 tcctgacata aagaatcgca tcgctaagcg cagcaggaag cttgtggatt atgatagtgc
     1021 tcgccatcac ctggaggctc tgcagagttc caaacggaaa gatgagagtc ggatctctaa
     1081 ggcagaagag gagtttcaga aagcacagaa agtgtttgaa gagttcaatg ttgacttaca
     1141 agaagagttg ccgtcattgt ggtcaagtcg agttggcttt tacgtcaata ctttcaaaaa
     1201 tgtctccagc cttgaggcca agtttcataa ggaaatcgca gtgctttgcc acaagctcta
     1261 tgaagtgatg acgaaactcg gtgatcagca tgctgacaag gccttcagca ttcaaggagc
     1321 tcccagtgac tcgggtcctc ttcgcattgc aaagacacca tcacctccag aggagccttc
     1381 tcccctgccc agccccacgg ctagtcccaa ccacacatta gcacctgcat ctcctgcccc
     1441 agtgcgaccc agatcacctt cacagacaag gaaagggcct cctgtcccac ctctgcctaa
     1501 agtcacccca acaaaggaac tgaaacagga gaacatcatc aatttctttg aggacaactt
     1561 tgtaccagaa atcaatgtga caacaccttc ccagaacgaa gtcttggagg tgaagaagga
     1621 agagactttg ctggatttgg actttgaccc tttcaagccc gatgtggctc cggcaggttc
     1681 tgctgcagcg acccactctc ccatgtctca gacattgccc tgggacttat ggacgacaag
     1741 cactgatttg gtacaaccag cttctggggg ttcatttaat gatttcaccc aggcccagga
     1801 cacttcatta ttcaccatgc agacagatca gaatatggct gaaactgagc aggctctacc
     1861 tacagagccc caagccgaag agcctcctgc cactgctgct gcacctactg ctgggctgga
     1921 ccttgggcta gagatggagg agccaaagga ggaggctgtg atccctcccg caactgatac
     1981 tggcgagact gtggaaactg cagtgccaac tgagggtgcc ccggtggagg aagcagaggc
     2041 agagaaggca gccctccctg ctggagaagg aggaagtcca gagggagcca aaattgacgg
     2101 ggaaagcacg gagcttgcaa tcagtgagag ccctcaacca gtggagcccg aagcgggtgc
     2161 tcctcaggtc atcccttctg ttgtcattga gccagcctcc aaccatgaag gagaaggaga
     2221 acaccaagaa actgctacgg gtactgaacc gagggaggcc gctgaggatg tggctgccca
     2281 gggctcagcg ggtgagaagc aggaagtggc tacagagccc acgcctctgg attctcaggc
     2341 aactctgcct gcatctgcag gtgctgtgga tgcctcgctg tctgcagggg atgccactca
     2401 ggaactgcct cctggctttc tctacaaggt ggaaaccctg catgatttcg aggcagcgaa
     2461 ctctgatgaa cttaacctac aaaggggtga cgtggtactc gtggtgcctt cagactcaga
     2521 agccgaccag gatgcaggct ggttggtggg ggtgaaggag tcagactggc ttcagtacag
     2581 agacctggcc acgtacaaag gcctctttcc agagaacttc acgcggcgcc tggagggaca
     2641 tggtacagga tcaactggta gtggaagtag tacccgggat ccaccggtcg ccaccatggt
     2701 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2761 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2821 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     2881 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     2941 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3001 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     3061 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     3121 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     3181 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     3241 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     3301 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3361 cgagggccgc cactccaccg gcggcatgga cgagctgtac aataaagcgg ccgcgactct
     3421 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3481 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3661 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3721 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3781 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3841 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3901 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3961 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4021 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4081 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4141 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4201 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4261 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4321 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4381 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4441 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4501 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4561 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4621 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4681 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4741 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4801 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4861 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4921 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4981 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5041 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5101 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5161 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5221 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5281 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5341 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5401 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5461 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5521 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5581 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5641 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5701 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5761 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5821 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5881 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5941 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6001 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6061 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6121 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6181 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6241 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6301 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6361 ggggttcgtg cacacagccc agcttkgagc gaacgaccta caccgaactg agatacctac
     6421 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6481 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6541 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6601 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6661 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6721 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     6781 agcccatata tggagttccg
//
