LOCUS       sequence-229623-        6703 bp    DNA     circular SYN 15-APR-2025
DEFINITION  GABAA receptor expression (chimeric rat delta subunit whose large 
            intracellular loop has been replaced with the large intracellular 
            loop of the rat gamma 2 short subunit) Myc-tag near N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  TITLE     de Blas Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Myc-delta/gamma2SIL
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..41)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(195..783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(275..294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(954..1745,1746..1814))
                     /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     1577..1596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1815..1919)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1994..2013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2185..2564
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2565..2768
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2719..2739
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2813..2832
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2813..2831
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2911..2940
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4244..4261)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4250..4474
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4520..4948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4607..4626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4817..4838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4957..4977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4962..5291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5142..5277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5204..5223
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5358..6152
                     /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(5412..5431)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6022..6041
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6326..6447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6363..6382)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6417..6436
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6496..6512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6496..6512)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6509..6531)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6520..6536
                     /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(6544..6550,6551..6568,6569..6574))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6589..6610
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
       61 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      601 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
      901 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
      961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1081 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1261 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1561 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1801 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1861 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1921 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     1981 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     2041 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     2101 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     2161 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     2221 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2281 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2341 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2401 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2461 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2521 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2581 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2641 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2701 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2761 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     2821 ctcactatag ggagacccaa gctggctagc atggacgttc tgggctggct gctgctgccg
     2881 ctccttctgc tgtgcacgca gccgcaccat gaacaaaaac taatatcaga agaagaccta
     2941 ggtgccagag caatgaatga cattggggac tacgtgggct ccaacctgga gatatcctgg
     3001 ctccccaacc tggatggact aatggagggc tacgcccgaa acttccgacc aggcattgga
     3061 ggtcctccag tgaatgtggc gcttgcccta gaggtggcca gcattgacca catctcagaa
     3121 gcaaatatgg aatacaccat gacagtgttc ctgcaccaga gctggcgaga cagcaggctg
     3181 tcctacaacc ataccaacga gaccctgggc ctggatagcc gcttcgtgga caagctgtgg
     3241 ctccctgaca ccttcattgt gaatgccaag tctgcctggt tccatgatgt gaccgtggaa
     3301 aacaagctta tccgcctaca gcccgacggt gtgattttat acagcatccg catcacctcc
     3361 acagtggcct gtgacatgga ccttgccaag taccccatgg acgagcagga gtgcatgctg
     3421 gacctggaga gctatggcta ctcttctgag gacattgtct attattggtc agaaaaccag
     3481 gagcagatcc acgggctgga caggctgcaa ctggcccagt tcactatcac cagttaccgc
     3541 ttcaccacgg agctgatgaa cttcaaatca gctggccagt tccctcgact cagcttacac
     3601 ttccagcttc ggaggaaccg gggtgtctac atcatccagt cttacatgcc ctctgtcctc
     3661 ctggttgcca tgtcctgggt ctccttctgg attagccaag cagcagtgcc tgccagagta
     3721 tctctaggca tcaccactgt gctgacaatg accacactca tggttagtgc ccgctcctcc
     3781 ctcccgcggg cttctgctat caaggctctg gatgtgtatt tctggatctg ctatgtcttc
     3841 gtgtttgctg ccctggtgga gtatgcattt gcccactatt ttgtgagcaa ccggaaacca
     3901 agcaaggata aagacaaaaa gaagaaaaac cctgccccta ccattgatat ccgtcccaga
     3961 tcagcaacga tccaaatgaa caatgccacc caccttcaag agagggatga agaatatggc
     4021 tatgagtgtt tggatggcaa ggactgtgcc agtttctttt gctgttttga agactgccga
     4081 acaggagcct ggagacacgg gaggatacac attcgcattg ccaaaatgga ctcctatgcc
     4141 cgcgctgtgt tcccggcagc ctttgcagca gtcaacatca tctactgggc agcgtatacc
     4201 atgtgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     4261 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4321 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4381 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4441 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4501 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4561 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4621 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4681 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4741 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4801 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4861 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4921 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4981 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5041 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5101 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5161 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5221 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5281 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5341 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5401 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5461 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5521 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5581 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5641 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5701 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5761 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5821 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5881 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5941 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6001 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6061 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6121 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     6181 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6241 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6301 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6361 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6421 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6481 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6541 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6601 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6661 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttg
//
