LOCUS       sequence-55792-e        7902 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 7902)
  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 7902)
  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(+)mGAT1-0-YFP
FEATURES             Location/Qualifiers
     source          1..7902
                     /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"
     regulatory      2741..2750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2747..2749,2750..2752,2753..3469)
                     /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
                     EFVTAAGITLGMDELYKI"
     primer_bind     complement(2792..2813)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3053..3072)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3400..3421
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3496..3513)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3502..3726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3772..4200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3859..3878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4069..4090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4209..4229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4214..4543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4394..4529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4456..4475
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4610..5404
                     /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(4664..4683)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5274..5293
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5578..5699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5615..5634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5669..5688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5748..5764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5748..5764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5761..5783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5772..5788
                     /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(5796..5802,5803..5820,5821..5826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5841..5862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5979..5996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6150..6735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6230..6249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6906..7697,7698..7766))
                     /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     7529..7548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7767..7871)
                     /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 tattgtgcgc cctgagaatg gccctgagca gccgcaggct ggcagctcag ccagcaagga
     2701 ggcctacatc cgaattctgc agatatccag cacagtggcg gccgccatgg tgagcaaggg
     2761 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2821 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2881 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     2941 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     3001 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3061 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3121 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3181 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3241 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3301 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     3361 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3421 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagatctaga gggcccgttt
     3481 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3541 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3601 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3661 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3721 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3781 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3841 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3901 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3961 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4021 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4081 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4141 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4201 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4321 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4381 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4441 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4501 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4561 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4621 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4681 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4741 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4801 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4861 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4921 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4981 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5041 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5101 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5161 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5221 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5281 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5341 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5401 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5461 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5521 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5701 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5761 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5821 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5881 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5941 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6661 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     6721 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6781 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6841 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6901 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6961 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7021 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7081 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7141 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7201 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7261 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7321 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7381 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7441 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7501 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7561 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7621 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7681 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7741 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7801 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7861 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
