LOCUS       Exported                4909 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To express GalT-mCherry in mammalian cells, GalT encodes the 1-81 
            amino acids from beta-1,4-galactosyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    pmCherry-N1-GalT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4909)
  AUTHORS   Tie HC, Mahajan D, Chen B, Cheng L, VanDongen AM, Lu L
  TITLE     A novel imaging method for quantitative Golgi localization reveals 
            differential intra-Golgi trafficking of secretory cargos.
  JOURNAL   Mol Biol Cell. 2016 Jan 13. pii: mbc.E15-09-0664.
  PUBMED    26764092
REFERENCE   2  (bases 1 to 4909)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4909
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     354..373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        628..931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        932..1135
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1086..1106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1425..1434
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1431..2141
                     /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(1576..1594)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1844..1863
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2264..2385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2301..2320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2355..2374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2392..2847)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2529..2550)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2741..2760
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2874..2978
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2980..3337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3003..3023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3188..3323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3250..3269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3372..4166
                     /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(3426..3445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4036..4055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4354..4373)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4398..4445
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4774..453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
       61 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      541 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
      601 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      661 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      721 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      781 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      841 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      901 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      961 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1021 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     1081 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     1141 agtgaaccgt cagatccgct agcatgaggc ttcgggagcc gctcctgagc ggcagcgccg
     1201 cgatgccagg cgcgtcccta cagcgggcct gccgcctgct cgtggccgtc tgcgctctgc
     1261 accttggcgt caccctcgtt tactacctgg ctggccgcga cctgagccgc ctgccccaac
     1321 tggtcggagt ctccacaccg ctgcagggcg gctcgaacag tgccgccgcc atcgggcagt
     1381 cctccgggga gctccggacc ggaggggcca aggatccacc ggtcgccacc atggtgagca
     1441 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     1501 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1561 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     1621 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     1681 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1741 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     1801 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     1861 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     1921 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     1981 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     2041 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     2101 gccgccactc caccggcggc atggacgagc tgtacaagta gagcggccgc gactctagat
     2161 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2221 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2281 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2341 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2401 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2461 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2521 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2581 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2641 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2701 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2761 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2821 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2881 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2941 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3001 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3061 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3181 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3241 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3301 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3361 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3421 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3481 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3541 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3601 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3661 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3721 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3781 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3841 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3901 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3961 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4021 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4081 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4141 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4201 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4261 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4321 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4381 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4441 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4501 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4561 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagata
//
