LOCUS       sequence-177296-        5171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GalT-iRFP670 in mammalian cells, GalT encodes the 1-81 
            amino acids from beta-1,4-galactosyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5171)
  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 5171)
  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: "Mol Biol 
            Cell. 2016 Jan 13. pii: mbc.E15-09-0664."
COMMENT     Sequence Label: piRFP670-N1-GalT
FEATURES             Location/Qualifiers
     source          1..5171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        64..421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        64..255
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(87..107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      272..407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             456..1250
                     /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             456..1250
                     /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(510..529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1120..1139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1438..1457)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1482..1529
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1858..2446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2347..2366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2621..2924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2925..3128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3079..3099
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3456..3465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3462..4397
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    4519..4640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4556..4575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4610..4629
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4647..5102)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4784..4805)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4996..5015
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5129..5171,1..62)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
       61 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
      121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      241 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      301 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
      361 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
      421 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
      481 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      541 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      601 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
      661 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      721 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      781 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      841 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      901 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
      961 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     1021 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1081 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1141 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1201 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     1261 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     1321 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1381 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     1441 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     1501 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     1561 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     1621 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     1681 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     1741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1801 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1861 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1921 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1981 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2041 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2101 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2161 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2221 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2281 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2341 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2401 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2461 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2521 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     2581 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2641 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2701 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2761 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2821 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2881 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2941 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3001 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     3061 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3121 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct
     3181 caagcttcga attcatgagg cttcgggagc cgctcctgag cggcagcgcc gcgatgccag
     3241 gcgcgtccct acagcgggcc tgccgcctgc tcgtggccgt ctgcgctctg caccttggcg
     3301 tcaccctcgt ttactacctg gctggccgcg acctgagccg cctgccccaa ctggtcggag
     3361 tctccacacc gctgcagggc ggctcgaaca gtgccgccgc catcgggcag tcctccgggg
     3421 agctccggac cggaggggcc gtggatccac cggtcgccac catggcgcgt aaggtcgatc
     3481 tcacctcctg cgatcgcgag ccgatccaca tccccggcag cattcagccg tgcggctgcc
     3541 tgctagcctg cgacgcgcag gcggtgcgga tcacgcgcat tacggaaaat gccggcgcgt
     3601 tctttggacg cgaaactccg cgggtcggtg agctactcgc cgattacttc ggcgagaccg
     3661 aagcccatgc gctgcgcaac gcactggcgc agtcctccga tccaaagcga ccggcgctga
     3721 tcttcggttg gcgcgacggc ctgaccggcc gcaccttcga catctcactg catcgccatg
     3781 acggtacatc gatcatcgag ttcgagcctg cggcggccga acaggccgac aatccgctgc
     3841 ggctgacgcg gcagatcatc gcgcgcacca aagaactgaa gtcgctcgaa gagatggccg
     3901 cacgggtgcc gcgctatctg caggcgatgc tcggctatca ccgcgtgatg ttgtaccgct
     3961 tcgcggacga cggctccggg atggtgatcg gcgaggcgaa gcgcagcgac ctcgagagct
     4021 ttctcggtca gcactttccg gcgtcgctgg tcccgcagca ggcgcggcta ctgtacttga
     4081 agaacgcgat ccgcgtggtc tcggattcgc gcggcatcag cagccggatc gtgcccgagc
     4141 acgacgcctc cggcgccgcg ctcgatctgt cgttcgcgca cctgcgcagc atctcgccct
     4201 gccatctcga atttctgcgg aacatgggcg tcagcgcctc gatgtcgctg tcgatcatca
     4261 ttgacggcac gctatgggga ttgatcatct gtcatcatta cgagccgcgt gccgtgccga
     4321 tggcgcagcg cgtcgcggcc gaaatgttcg ccgacttctt atcgctgcac ttcaccgccg
     4381 cccaccacca acgctaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     4441 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4501 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4621 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4681 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4741 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4801 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4861 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4921 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4981 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5041 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5161 acattcaaat a
//
