LOCUS       sequence-388452-        6256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of an epitope-tagged hOLIG1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6256)
  TITLE     Trimmer lab antigen plasmids
REFERENCE   2  (bases 1 to 6256)
  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"
COMMENT     Sequence Label: hOlig1-myc/pCDNA3.1
FEATURES             Location/Qualifiers
     source          1..6256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             950..979
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             995..1012
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1035..1052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1041..1265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1311..1739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1398..1417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1608..1629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1748..1768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1753..2082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1933..2068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1995..2014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2149..2943
                     /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(2203..2222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2813..2832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3119..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3156..3175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3210..3229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3289..3305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3302..3324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3313..3329
                     /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(3337..3343,3344..3361,3362..3367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3382..3403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3520..3537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3771..3790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4450..5241,5242..5310))
                     /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     5073..5092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5311..5415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5490..5509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5681..6060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6061..6256,1..8)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6215..6235
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
       61 ctcactatag ggagacccaa gctggctagt taagcttaat atgctgcggc cacagcggcc
      121 cggagacttg cagctcgggg cctccctcta cgagctggtg ggctacaggc agccgccctc
      181 ctcctcctcc tcctccacct cctccacctc ctccacttcc tcctcctcca cgacggcccc
      241 cctcctcccc aaggctgcgc gcgagaagcc ggaggcgccg gccgagcctc caggccccgg
      301 gcccgggtca ggcgcgcacc cgggcggcag cgcccggccg gacgccaagg aggagcagca
      361 gcagcagctg cggcgcaaga tcaacagccg cgagcggaag cgcatgcagg acctgaacct
      421 ggccatggac gccctgcgcg aggtcatcct gccctactca gcggcgcact gccagggcgc
      481 gcccggccgc aagctctcca agatagccac gctgctgctc gcccgcaact acatcctact
      541 gctgggcagc tcgctgcagg agctgcgccg cgcgctgggc gagggcgccg ggcccgccgc
      601 gccgcgcctg ctgctggccg ggctgcccct gctcgccgcc gcgcccggct ccgtgttgct
      661 ggcgcccggc gccgtaggac cccccgacgc gctgcgcccc gccaagtacc tgtcgctggc
      721 gctggacgag ccgccgtgcg gccagttcgc tctccccggc ggcggcgcag gcggccccgg
      781 cctctgcacc tgcgccgtgt gcaagttccc gcacctggtc ccggccagcc tgggcctggc
      841 cgccgtgcag gcgcaattct ccaagaatgg atccactagt ccagtgtggt ggaattctgc
      901 agatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg aacaaaaact
      961 catctcagaa gaggatctga atatgcatac cggtcatcat caccatcacc attgagttta
     1021 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1081 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1141 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1201 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1261 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1381 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1501 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1621 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1681 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     1741 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     1801 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     1861 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     1921 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     1981 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2041 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2101 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2161 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2221 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2281 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     2341 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2401 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2461 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2521 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2581 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2641 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     2701 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2761 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2821 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2881 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2941 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3001 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3061 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3241 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     5461 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5521 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     5581 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     5641 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     5701 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5761 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5821 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5881 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5941 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6001 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6061 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     6121 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6181 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     6241 tgggaggtct atataa
//
