LOCUS       sequence-490927-        6754 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  fusion of Rab5c to mTFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Vigano MA, Bieli D, Schaefer JV, Jakob RP, Matsuda S, Maier T, 
            Pluckthun A, Affolter M
  TITLE     DARPins recognizing mTFP1 as novel reagents for in vitro and in vivo
            protein manipulations.
  JOURNAL   Biol Open. 2018 Oct 29;7(11). pii: bio.036749. doi: 
            10.1242/bio.036749.
  PUBMED    30237292
REFERENCE   2  (bases 1 to 6754)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biol 
            Open."; date: "2018-10-29"; volume: "7(11). pii: bio.036749. doi"; 
            pages: " 10.1242/bio.036749"
COMMENT     Sequence Label: pcDNA3-mTFP1-Rab5c
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             902..1609
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     promoter        complement(2307..2325)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2308..2325)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2345..2362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2351..2575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2621..3049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2708..2727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2918..2939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3058..3078)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3063..3392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3243..3378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3305..3324
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3459..4253
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3513..3532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4123..4142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4427..4548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4464..4483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4518..4537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4597..4613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4597..4613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4610..4632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4621..4637
                     /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(4645..4651,4652..4669,4670..4675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4690..4711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4828..4845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4999..5587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5079..5098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5758..6549,6550..6618))
                     /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     6381..6400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6619..6723)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 catggtgagc aagggcgagg agaccacaat gggcgtaatc aagcccgaca tgaagatcaa
      961 gctgaagatg gagggcaacg tgaatggcca cgccttcgtg atcgagggcg agggcgaggg
     1021 caagccctac gacggcacca acaccatcaa cctggaggtg aaggagggag cccccctgcc
     1081 cttctcctac gacattctga ccaccgcgtt cgcctacggc aacagggcct tcaccaagta
     1141 ccccgacgac atccccaact acttcaagca gtccttcccc gagggctact cttgggagcg
     1201 caccatgacc ttcgaggaca agggcatcgt gaaggtgaag tccgacatct ccatggagga
     1261 ggactccttc atctacgaga tacacctcaa gggcgagaac ttccccccca acggccccgt
     1321 gatgcagaag aagaccaccg gctgggacgc ctccaccgag aggatgtacg tgcgcgacgg
     1381 cgtgctgaag ggcgacgtca agcacaagct gctgctggag ggcggcggcc accaccgcgt
     1441 tgacttcaag accatctaca gggccaagaa ggcggtgaag ctgcccgact atcactttgt
     1501 ggaccaccgc atcgagatcc tgaaccacga caaggactac aacaaggtga ccgtttacga
     1561 gagcgccgtg gcccgcaact ccaccgacgg catggacgag ctgtacaaag tggtcaccat
     1621 ggcggggcga ggtggaccag cgcggaccaa cggtgccgcc cccgtcggga acaaaatctg
     1681 ccagtttaag cttgtgttgc tgggagagtc tgcagtaggc aagtccagcc tggtgctgcg
     1741 cttcgtcaaa ggccagtttc acgagtacca ggagagcacc attggagctg ctttcctcac
     1801 acagacactc tgcttggacg atacgacagt aaagtttgag atttgggaca cggccggaca
     1861 ggagcggtat cacagtttgg cccctatgta ttacagagga gcccaggcag ccatcgtcgt
     1921 ctacgacatt actaacacag acacattcac tagagcaaag aactgggtga aggagctgca
     1981 gcgccaggcc agtccaaata tcgttattgc tctggcaggc aacaaagcag atctggccaa
     2041 caagagagcc gttgactttc aggaggccca ggcatacgca gatgacaaca gcctactctt
     2101 catggagacg tctgcaaaga ccgccatgaa cgtcaatgag atcttcatgg ctatcgccaa
     2161 gaagcttccg aaaagcgagc cgcagggtgg agcgggcagc gggggacggg cccgaggagg
     2221 ggtggactta caggagacgg cgcctcaggg aagaagcggc cagtgctgtg gaggcggaaa
     2281 ctaaggaatt ctctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     2341 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2401 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2461 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2521 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2581 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2761 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2821 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2941 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3001 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3061 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3301 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3361 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3421 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6721 gcgcacattt ccccgaaaag tgccacctga cgtc
//
