LOCUS       sequence-55832-e        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  AUTHORS   Moss FJ, Imoukhuede PI, Scott K, Hu J, Jankowsky JL, Quick MW, 
            Lester HA
  TITLE     GABA transporter function, oligomerization state, and anchoring: 
            correlates with subcellularly resolved FRET.
  JOURNAL   J Gen Physiol. 2009 Dec;134(6):489-521.
  PUBMED    19948998
REFERENCE   2  (bases 1 to 7906)
  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: "J Gen 
            Physiol."; date: "2009-12"; volume: "134(6)"; pages: "489-521"
COMMENT     Sequence Label: pcDNA3.1(+)mGAT1-577-YFP-578CT
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2642..2644,2645..2647,2648..3358)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2687..2708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2948..2967)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3295..3316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3500..3517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3506..3730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3776..4204
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3863..3882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4073..4094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4213..4233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4218..4547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4398..4533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4460..4479
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4614..5408
                     /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(4668..4687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5278..5297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5582..5703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5619..5638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5673..5692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5752..5768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5752..5768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5765..5787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5776..5792
                     /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(5800..5806,5807..5824,5825..5830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5845..5866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5983..6000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6154..6739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6234..6253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6910..7701,7702..7770))
                     /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     7533..7552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7771..7875)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgg cgactgacaa cagcaaggtg gctgatgggc agatctctac
      961 tgaggtcagc gaggcccctg tggccagcga caagcccaaa accctggtag tcaaggtgca
     1021 gaagaaggcc ggggacctcc ctgaccggga cacatggaag ggacgcttcg acttcctcat
     1081 gtcctgcgtg ggctatgcca tcggcctggg caatgtgtgg aggttccctt acctctgtgg
     1141 gaaaaacggt ggcggggcct tcctaatccc atatttcctg acgctcatct ttgcgggtgt
     1201 tcctctcttc cttttggagt gctccctagg ccagtacacc tccattgggg gcctgggcgt
     1261 atggaactgg gcgcccatgt tcaagggtgt ggccgtcgcg gcagctgtgc tgtccttctg
     1321 gctgaacatc tactacatcg tcatcatctc ctgggccatc tactacctgt acaactcctt
     1381 caccacgacc ctgccatgga aacagtgtga caacccgtgg aacactgacc gctgcttctc
     1441 caactacagc ctggtcaata ccaccaacat gaccagcgcc gtggtggagt tctgggagcg
     1501 caacatgcac cagatgacag atggactgga caagccagga cagatccgct gtctggccat
     1561 cacactggcc attgcctggg tgctcgtgta tttctgcatc tggaagggtg ttggttggac
     1621 tggaaaggtg gtctacttct cagccacgta cccctacatc atgcttatca tcctgttctt
     1681 ccgtggagtg acgcttcccg gggccaagga ggggatcctc ttctacatca cacccaactt
     1741 ccgaaagctg tctgattctg aggtgatctt tgacgccgcc acccagatct tcttctccta
     1801 cgggctgggc ctggggtccc tgattgctct gggaagctac aactctttcc acaacaatgt
     1861 gtacagggac tccatcatcg tttgctgcat caactcctgc accagcatgt ttgccggatt
     1921 cgtcatcttc tccatcgtgg gcttcatggc tcatgtcacc aagaggtcca tagctgatgt
     1981 ggcagcctca ggcccggggc tggcattctt ggcgtaccct gaggctgtga cacagctacc
     2041 catctctccc ctctgggcta tcctcttctt ctccatgctg ctgatgctgg gcattgacag
     2101 ccagttctgt accgtggagg gcttcatcac tgccctggtg gacgagtacc ccagacttct
     2161 ccgcaatcgc cgtgaactct tcattgctgc cgtgtgcatc gtgtcctacc tgattggcct
     2221 gtctaacatc acccagggtg gcatttatgt cttcaaactg tttgattatt actctgccag
     2281 cggcatgagc ttgctgttcc tggttttctt cgagtgtgtc tccatttcct ggttttatgg
     2341 tgtcaaccgg ttctatgaca acatccagga gatggttggc tccaggccct gcatctggtg
     2401 gaagctgtgc tggtcctttt tcacacccat cattgtggcg ggcgtgtttc tcttcagtgc
     2461 tgtgcagatg acaccactca ccatgggaag ctatgttttc cccaagtggg gccagggcgt
     2521 gggctggctc atggctctgt cctccatggt gctcatcccc gggtacatgg cttacatgtt
     2581 cctcaccctg aagggctccc tgaagcagcg tctccaggtc atgattcagc ccagtgaaga
     2641 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2701 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2761 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2821 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     2881 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2941 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3001 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3061 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3121 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3181 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3241 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     3301 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagat
     3361 tgtgcgccct gagaatggcc ctgagcagcc gcaggctggc agctcagcca gcaaggaggc
     3421 ctacatctag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     3481 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3541 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3601 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3661 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3721 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3901 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4081 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4141 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4201 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4321 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4381 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4441 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4501 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4561 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4621 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4681 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4741 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4801 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4861 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4921 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4981 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5041 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5101 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5161 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5221 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5281 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5341 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5401 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
