LOCUS       sequence-193958-        7145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Glt1b delta 4 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  TITLE     Rosenberg lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7145)
  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            "Unpublished"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: GLT-1b delta 4
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2464..2482)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2465..2482)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2502..2519)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2508..2732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2778..3206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2865..2884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3075..3096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3215..3235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3220..3549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3400..3535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3462..3481
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3616..4410
                     /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(3670..3689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4280..4299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4584..4705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4621..4640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4675..4694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4754..4770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4754..4770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4767..4789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4778..4794
                     /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(4802..4808,4809..4826,4827..4832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4847..4868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4985..5002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5156..5744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5236..5255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5915..6706,6707..6775))
                     /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     6538..6557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6776..6880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6955..6974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg cccttcgcca
      721 tggcatcaac cgagggtgcc aacaatatgc ccaagcaggt ggaagtgcgc atgcacgaca
      781 gccacctcag ctccgaggag ccaaagcacc gaaacctggg catgcgcatg tgcgacaagc
      841 tggggaagaa cctcctgctc tcactgactg tgtttggtgt catcctggga gcagtatgtg
      901 gcgggctgct tcgcttggcg gctcccatcc accctgatgt ggtcatgttg atagccttcc
      961 cgggggacat cctcatgagg atgctgaaga tgctcatcct ccctctcatc atttccagtc
     1021 tcatcacagg gctgtcaggg ctggatgcta aagccagcgg ccgcctaggc acgagagcca
     1081 tggtatatta catgtccacg accatcattg ccgctgtgct gggggtcatc ctggtgttgg
     1141 ccatccaccc tgggaatccc aaactcaaga agcagctggg gcctgggaag aagaacgacg
     1201 aggtgtccag cctggatgcc ttcctggatc tcattagaaa tctcttcccg gagaacctgg
     1261 tacaagcctg tttccaacag attcagactg tgacaaagaa agttctggtg gcacctccat
     1321 ccgaggaggc caatacaacc aaggcagtca tctccctgtt gaatgagacc atgaatgagg
     1381 cccctgaaga aactaagatc gttatcaaga agggcctgga gttcaaggac gggatgaatg
     1441 tcttaggtct gattggattc tttattgctt tcggcattgc catggggaag atgggtgagc
     1501 aggccaagct gatggtggag ttcttcaaca ttctgaacga gattgtcatg aagttagtga
     1561 tcatgatcat gtggtattcc ccgctgggta tcgcctgctt gatctgtggg aagatcatcg
     1621 ccatcaagga cttagaagtg gttgctaggc agctggggat gtacatgatc acagtgatcg
     1681 tgggcctcat cattcatggg ggcatctttc tccccttgat ttactttgta gtgaccagaa
     1741 aaaacccatt ctcctttttt gctggcattt tccaagcctg gatcactgcc ctgggaaccg
     1801 cttccagtgc tggaactttg cctgtcacct tccgttgctt ggaagataat ctagggattg
     1861 acaagcgtgt gaccagattc gtcctcccag tcggagcaac cattaacatg gatggtacag
     1921 ccctttacga agccgtggca gccatcttca tagcccagat gaatggggtc atcctggatg
     1981 gaggtcagat agtgactgta agccttacag caactctggc gagcattggt gcagccagta
     2041 ttcccagcgc cgggttggtc accatgctcc tcattctcac agctgtgggc ctgccgacag
     2101 aggacatcag tctgctggtg gcagtggact ggctgctgga tagaatgaga acttcggtca
     2161 atgtagtggg cgattctttt ggggctggga ttgtctatca cctttccaag tccgagctgg
     2221 acaccattga ctcccaacac cgaatgcacg aagacatcga aatgaccaag acgcagtccg
     2281 tttatgacga cacgaagaac cacagggaaa gcaactctaa tcagtgtgtc tatgccgcac
     2341 acaactctgt cgtaatagat gagtgcaagg tacctttccc attcctggat atctgataag
     2401 ggcgaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     2461 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     2521 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2581 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2641 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2701 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2761 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2821 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2881 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2941 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3001 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3061 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3121 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3181 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3481 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3541 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3601 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3661 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3721 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3781 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3841 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3901 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3961 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4021 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4081 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4141 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4201 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4261 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4321 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4381 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4441 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4501 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4561 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4681 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4741 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4861 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5821 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5881 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5941 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6001 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6061 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6121 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6181 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6241 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6301 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6361 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6421 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6481 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6541 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6601 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6661 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6721 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6781 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6841 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6901 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     6961 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7021 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7081 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     7141 gcgtt
//
