LOCUS       sequence-9433-ex        7683 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 7683)
  AUTHORS   Saunders C, Ferrer JV, Shi L, Chen J, Merrill G, Lamb ME, 
            Leeb-Lundberg LM, Carvelli L, Javitch JA, Galli A
  TITLE     Amphetamine-induced loss of human dopamine transporter activity: an 
            internalization-dependent and cocaine-sensitive mechanism.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 Jun 6. 97(12):6850-5.
  PUBMED    10823899
REFERENCE   2  (bases 1 to 7683)
  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: "Proc Natl 
            Acad Sci U S A. 2000 Jun 6. 97(12):6850-5."
COMMENT     Sequence Label: pcihygroflagsynhDAT
FEATURES             Location/Qualifiers
     source          1..7683
                     /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      926..935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          2926..3155
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3138..3157
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3231..3806,3807..3809,3810..3818)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3398..3415)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(5170..5187)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5176..5400
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5447..5466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5526..5542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5526..5542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5539..5561)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5550..5566
                     /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(5574..5580,5581..5598,5599..5604))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5619..5640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5757..5774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5928..6516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6008..6027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6687..7478,7479..7547))
                     /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     7310..7329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7548..7652)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 ccgggtgtac aaggttaacg gtaccgccac catggactac aaggacgatg acgataaggc
      961 cagcaagtcc aaatgcagcg tgggactgat gtccagcgtg gttgcccccg ctaaggagcc
     1021 caacgccgtg ggccccaagg aggtggaact gattctcgtg aaggagcaga acggcgtgca
     1081 gctcacctcc agcacactga ccaaccctag gcagagcccc gtggaggccc aggaccggga
     1141 gacctggggc aagaaaatcg acttcctgct cagcgtgatc ggcttcgccg tcgacctggc
     1201 taacgtttgg cggttcccct acctgtgcta caagaacggc ggaggcgctt tcctggtgcc
     1261 ctacctcctc tttatggtga tcgctggcat gcccctgttc tacatggagc tggccctcgg
     1321 ccagtttaac cgggagggag ccgcaggtgt gtggaagatc tgccccatcc tcaaaggcgt
     1381 gggattcacc gtgatcctga tcagcctgta cgtaggcttc ttttacaacg tcatcattgc
     1441 ctgggctctc cactacctgt tctcgagctt caccacagag ctgccctgga tccactgcaa
     1501 caatagctgg aacagcccca actgcagcga tgcgcacccc ggcgacagct ccggagacag
     1561 ctcaggcctg aacgacacct tcggaaccac acccgccgct gaatacttcg aacggggcgt
     1621 gctgcacctc catcagagcc acggcatcga tgaccttggt cctcccaggt ggcagctgac
     1681 cgcctgtctg gtgctcgtca tcgtgctgct ctacttcagc ctgtggaagg gcgtgaagac
     1741 aagcggcaag gtggtctgga tcaccgccac aatgccatat gtggtcctga ccgccctgct
     1801 cctgcggggg gtgaccctgc ctggggccat cgacggcatt cgggcctacc tgagcgtgga
     1861 cttctaccgg ctgtgcgagg ctagcgtgtg gatcgacgct gccactcaag tgtgcttcag
     1921 cctgggcgtg ggattcgggg tcctgatcgc cttcagctcc tacaacaagt tcaccaacaa
     1981 ttgctaccgc gacgccatcg tcaccacaag catcaactcc ctgactagtt tttcgagcgg
     2041 ctttgtcgtg ttcagctttc tgggatacat ggcccagaag cacagcgtgc ccatcggtga
     2101 cgtcgccaag gacggccccg ggctgatctt cattatctac cctgaggcca tcgctaccct
     2161 gcccctgagc tccgcctggg ctgtcgtgtt ctttatcatg ctgctcaccc tcggcatcga
     2221 cagcgctatg ggaggcatgg agagcgtcat caccggcctg atcgacgaat tccagctcct
     2281 gcaccggcat cgcgagctgt tcaccctgtt catcgtgctg gccaccttcc ttctgagcct
     2341 gttctgcgtg accaacggcg gaatctacgt gttcaccctc ctggaccact tcgctgccgg
     2401 cacctcaata ttgttcggcg tgctgatcga ggcgatcggc gtggcctggt tctacggagt
     2461 gggccagttc agcgacgata tccagcagat gaccggtcag cggcccagcc tgtactggcg
     2521 gctgtgctgg aagctcgtct ccccctgctt cctcctgttc gtggtcgttg tgtcaattgt
     2581 gaccttccgg cctccccact acggcgccta catcttcccc gactgggcca atgcattggg
     2641 ctgggtgatc gcaaccagct ccatggctat ggtgcccatc tacgctgcct acaagttctg
     2701 tagcctgccc ggaagcttcc gggagaagct ggcgtacgct atcgcccccg agaaggaccg
     2761 ggagctggtg gaccgcggcg aggtgcggca gttcaccctg cggcactggc tgaaggtgtg
     2821 aagagagtct agaagcggcc gcgtcgacgg aattcagtgg atccactagt aacggccgcc
     2881 agtgtgctgg aattaattcg ctgtctgcga gggccggctg ttggggtgag tactccctct
     2941 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     3001 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     3061 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     3121 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     3181 ggtcgatcga gcatgcatct agggcggccg gccgcactag aggaattccg cccctctccc
     3241 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgtgtttgtc
     3301 tatatgtgat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     3361 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     3421 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     3481 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     3541 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     3601 tggatagttg tggaaagagt caaatggctc tcctcaagcg tagtcaacaa ggggctgaag
     3661 gatgcccaga aggtacccca ttgtatggga atctgatctg gggcctcggt gcacatgctt
     3721 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     3781 tttcctttga aaaacacgat gataagcttg ccacaacccg taccaaagat ggatagatcc
     3841 ggaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     3901 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     3961 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     4021 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     4081 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     4141 gacctgcctg aaaccgaact gcccgctgtt ctgcaccggt cgcggaggcc atggatgcga
     4201 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4261 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4321 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     4381 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     4441 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     4501 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     4561 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     4621 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     4681 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     4741 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4801 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4861 agagtagatg ccgaccgaac aagagctgat ttcgagaacg cctcagccag caactcgcgc
     4921 gagcctagca aggcaaatgc gagagaacgg ccttacgctt ggtggcacag ttctcgtcca
     4981 cagttcgcta agctcgctcg gctgggtcgc gggagggccg gtcgcagtga ttcaggccct
     5041 tctggattgt gttggtcccc agggcacgat tgtcatgccc acgcactcgg gtgatctgac
     5101 tgatcccgca gattggagat cgccgcccgt gcctgccgat tgggtgcaga tctagagctc
     5161 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5221 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5281 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5341 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5401 cttctgaggc ggaaagaacc agctggggct cgagtgcatt ctagttgtgg tttgtccaaa
     5461 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5521 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5581 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5641 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5701 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6121 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7681 gtc
//
