LOCUS       sequence-55829-e        7908 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 7908)
  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 7908)
  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(+)mGAT1-570-YFP-571CT
FEATURES             Location/Qualifiers
     source          1..7908
                     /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(2627..2629,2630..2632,2633..3343)
                     /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(2672..2693)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2933..2952)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3280..3301
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3502..3519)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3508..3732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3778..4206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3865..3884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4075..4096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4215..4235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4220..4549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4400..4535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4462..4481
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4616..5410
                     /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(4670..4689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5280..5299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5584..5705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5621..5640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5675..5694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5754..5770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5754..5770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5767..5789)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5778..5794
                     /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(5802..5808,5809..5826,5827..5832))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5847..5868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5985..6002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6156..6741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6236..6255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6912..7703,7704..7772))
                     /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     7535..7554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7773..7877)
                     /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 aggtgtggct 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 atgattatgg tgagcaaggg
     2641 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2701 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2761 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     2821 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2881 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2941 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3001 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3061 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3121 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3181 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     3241 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3301 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagcagccca gtgaagatat
     3361 tgtgcgccct gagaatggcc ctgagcagcc gcaggctggc agctcagcca gcaaggaggc
     3421 ctacatctag cgaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc
     3481 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3541 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3601 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3661 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3721 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3781 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3841 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3901 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3961 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4021 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4081 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4141 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4201 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4261 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4381 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4441 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4501 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4561 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4621 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4681 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4741 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4801 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4861 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4921 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4981 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5041 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5101 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5161 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5221 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5281 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5341 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5401 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5461 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5521 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5581 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5701 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5761 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5821 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5881 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     6721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6841 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6901 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6961 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7021 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7081 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7141 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7201 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7261 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7321 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7381 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7441 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7501 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7561 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7621 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7681 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7741 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7801 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
