LOCUS       sequence-55819-e        7983 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 7983)
  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 7983)
  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(+)mGAT1YFP28
FEATURES             Location/Qualifiers
     source          1..7983
                     /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..3463)
                     /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(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(3577..3594)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3583..3807
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3853..4281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3940..3959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4150..4171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4290..4310)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4295..4624
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4475..4610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4537..4556
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4691..5485
                     /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(4745..4764)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5355..5374
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5659..5780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5696..5715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5750..5769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5829..5845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5829..5845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5842..5864)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5853..5869
                     /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(5877..5883,5884..5901,5902..5907))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5922..5943
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6060..6077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6231..6816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6311..6330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6987..7778,7779..7847))
                     /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     7610..7629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7848..7952)
                     /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 aagcagccca gcgaagacat
     3481 cgttcgccca gagaatggtc ctgagcagcc ccaggcgggc agctccacca gcaaggaggc
     3541 ctacatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3601 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3661 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3721 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3781 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3841 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3901 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3961 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4021 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4081 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4141 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4201 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4261 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4381 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4441 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4501 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4561 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4621 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4681 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4741 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4801 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4861 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4921 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4981 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5041 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5101 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5161 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5221 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5281 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5341 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5401 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5461 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5521 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5581 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5641 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5701 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5761 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5821 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5881 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5941 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     6781 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6841 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6901 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6961 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7021 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7081 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7141 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7201 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7261 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7321 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7381 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7441 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7501 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7561 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7621 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7681 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7741 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7801 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7861 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7921 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7981 gtc
//
