LOCUS       sequence-443045-        6694 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 6694)
  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 6694)
  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: CALM-pmCherryN1
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(321..425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     749..770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    914..1035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     979..998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1163..1867)
                     /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(1435..1454)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1704..1722
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1864..1873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3949..4152)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3978..3998)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4153..4456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4631..5219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4711..4730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5548..5595
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5620..5639
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5827..6621)
                     /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             complement(5827..6621)
                     /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(5938..5957)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6548..6567
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6656..6694,1..319))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6670..6694,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
      361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      901 acaatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1141 ctagagtcgc ggccgcttta ttgtacagct cgtccatgcc gccggtggag tggcggccct
     1201 cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccaact
     1261 tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct
     1321 tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct
     1381 tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct
     1441 tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga
     1501 actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt
     1561 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt
     1621 cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc
     1681 aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     1741 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca
     1801 tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc tcgcccttgc
     1861 tcaccatggt ggcgaccggt ggatcccggg ccagctgacc cttggtcgaa cccgaggtcg
     1921 aacccatccg cataaactgt atctgtgctc ctgatacagg gccaaagggg tttggtggtc
     1981 tcatgacagg ctggctgtat attaaggttg gttgtgtcat tacaggtaca cttcccatct
     2041 gaggaggaat tccatatcct atcatgcccg ttggtgtagt agcaggatag gccattacag
     2101 ggggcgcgta ttgtggaaaa tgcatgccat tcattgttgc agcgctccag gcagttgttg
     2161 gtgcaacctt tggttgccag ttagatcctc cagttaactt cttttcacct ggttgactcc
     2221 aacttacatc attcttagtg gttccatttc caatgccaag attgcccaca aggttggcta
     2281 aagatgaatc caagtcgtca gacactaatt tgttaggcgg aagtttggca acaggaagac
     2341 tctggttctg ggaggccact gttggtttga gaagtccacc accagaatct acagcaacat
     2401 tcgtagattt atttccaaac acagattcaa agtcaacatt aaggccagct gaagaatgtg
     2461 gctgtgcaac tggagaagga ctgaatccaa caaacatctc atgagtaggt gtcctagtag
     2521 taaaagtgga tacatctgaa gcaatgggca ggtgaacatc accactactt tttgtgagga
     2581 aagggtttaa gcttggaatg gcatcctcaa cagcatctag agtagcagag aaaggatctc
     2641 cccatgtact cgctccctga ggagcagccg acatggcgtg gacagacggg tgaaaggttg
     2701 gctgctgcaa gtcaagcagg tcatttggca gcttagatgt gctgttagaa gaactagggg
     2761 tagaaaatat gtcgatggct ggtgcagtca ttattccccc tgctgaggtg gacaccggag
     2821 aggctgcagt tgttagagag gtatgcggtt tctttgcaag ttcttttaga cgctgttcct
     2881 ttaatgcttt taaacgagcc tgttcttcct ctaatgctgc ctgtttttcc ctttcatcca
     2941 ctttggtcag agataggcca gtgcttgcca aagaagagac tgcattggaa agtgttgtag
     3001 ccctgcttgc agctgtggaa tctttgattt tctttccttc caaggaagct aaatgttgtt
     3061 ctaaagcatc aagaagactg ctgggggcct gtgaaagatc tggaatatct cctctgtcaa
     3121 ttccaacttg ctctgcaact ttcagaaact ctgagattct tgtcatccta gtcagaaact
     3181 tcttatagat gtcaagacct tctttgcact ggttcttttt catatcaaaa tatttttcca
     3241 ataaattaat aattccttca ttgtatgctg caaatagtct aatggcatct ttgaagagga
     3301 gcatgaaggc agcatttatt accccatttg taagttcatt actattaaca ttaaaatcaa
     3361 gaagtgcatc catttgattt tggataattg gtacagtttt taagagtttt tctgtgttca
     3421 ttgttctcat aactccatca gctcctctct tcacttttgt gaaatcgaat gcaacttgtc
     3481 tgtatgaaac tgctttttca tttaagtatc tactataccg tctaataaat gtagacatat
     3541 catatccttg caatccactt ttatccaaaa aattgcttaa gttaaacaat gtgtttcttg
     3601 aagccaaata ctgaatgaaa cgctcatttc cataaaccat caaatgatga gttgtaatga
     3661 gtgatttgaa gaccaccacc caactactgt tagtagttct ttcaaacaaa ctgtccgcca
     3721 actgtgggat gttcacattc atctcatttg tacactgaat taagtagtcc aggtgctttt
     3781 tcttggggcc catgatctcg tgggtcgtgg ccttgcatac tgtcttagat accgcggagc
     3841 cagtgacact gtgctgggcc gcggtgattc ggtccgtcag gctctggcca gacatggtct
     3901 cgagatctga gtccggtagc gctagcggat ctgacggttc actaaaccag ctctgcttat
     3961 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     4021 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     4081 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     4141 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     4201 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     4261 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     4321 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     4381 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     4441 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     4501 ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca ggggataacg
     4561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4621 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5341 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     5401 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     5461 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     5521 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     5581 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     5641 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     5701 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     5761 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     5821 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     5881 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     5941 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     6001 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     6061 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     6121 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     6181 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     6241 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     6301 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     6361 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     6421 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     6481 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     6541 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     6601 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc
     6661 aaaagcctag gcctccaaaa aagcctcctc acta
//
