LOCUS       sequence-135343-        4811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo protein protein interaction studies/ expression in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4811)
  TITLE     pGolt1-mCherry, expression in mammalian cell
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4811)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGolt1-mCherry
FEATURES             Location/Qualifiers
     source          1..4811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      760..769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             766..1476
                     /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(911..929)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1179..1198
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1599..1720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1636..1655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1690..1709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1727..2182)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1864..1885)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2076..2095
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2209..2313
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2315..2672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2315..2506
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2338..2358)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2523..2658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2707..3501
                     /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             2707..3501
                     /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(2761..2780)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3371..3390
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3689..3708)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3733..3780
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatatgagcc tcatctgcgc cttcgccatc gccttcatca tcagtgtcat gctcatcgcg
      721 gccaaccagg tgctgcgcag tggcatgaag ccaccggtcg ccaccatggt gagcaagggc
      781 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
      841 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      901 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
      961 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1021 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1081 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1141 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     1201 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     1261 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     1321 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     1381 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     1441 cactccaccg gcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     1501 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1561 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1681 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1741 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1801 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1861 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1921 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1981 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2041 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2101 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2161 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2221 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2281 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2341 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2401 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2461 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2521 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2581 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2641 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2701 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2761 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2821 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2881 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2941 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3001 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3061 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3121 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3181 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3241 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3301 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3361 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3421 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3481 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3541 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3601 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3661 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3721 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3781 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3841 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3901 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     3961 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4021 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4801 ccgccatgca t
//
