LOCUS       sequence-55812-e        8034 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 8034)
  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 8034)
  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(+)mGAT1YFP45
FEATURES             Location/Qualifiers
     source          1..8034
                     /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(3628..3645)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3634..3858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3904..4332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3991..4010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4201..4222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4341..4361)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4346..4675
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4526..4661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4588..4607
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4742..5536
                     /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(4796..4815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5406..5425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5710..5831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5747..5766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5801..5820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5880..5896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5880..5896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5893..5915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5904..5920
                     /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(5928..5934,5935..5952,5953..5958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5973..5994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6111..6128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6282..6867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6362..6381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7038..7829,7830..7898))
                     /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     7661..7680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7899..8003)
                     /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 aagatgttcc tcaccttaaa
     3481 gggctccctg aagcagcgca tccaagtcat ggtccagccc agcgaagaca tcgttcgccc
     3541 agagaatggt cctgagcagc cccaggcggg cagctccacc agcaaggagg cctacatcta
     3601 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3661 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3721 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3781 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3841 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3901 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3961 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4021 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4081 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4141 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4201 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4261 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4321 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4381 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4501 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4561 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4621 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4681 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4741 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4801 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4861 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4921 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4981 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5041 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5101 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5161 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5221 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5281 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5341 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5401 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5461 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5521 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5581 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5641 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5701 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5761 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5821 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5881 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5941 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6001 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6061 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6181 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     6841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
