LOCUS       sequence-9435-ex        7624 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 7624)
  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 7624)
  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: pcihygrosynDAT
FEATURES             Location/Qualifiers
     source          1..7624
                     /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"
     intron          2871..3100
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3083..3102
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3172..3747,3748..3750,3751..3759)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3339..3356)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(5111..5128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5117..5341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5388..5407
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5467..5483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5467..5483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5480..5502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5491..5507
                     /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(5515..5521,5522..5539,5540..5545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5560..5581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5698..5715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5869..6457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5949..5968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6628..7419,7420..7488))
                     /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     7251..7270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7489..7593)
                     /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 accatgagca agtccaaatg cagcgtggga ctgatgtcca gcgtggttgc ccccgctaag
      961 gagcccaacg ccgtgggccc caaggaggtg gaactgattc tcgtgaagga gcagaacggc
     1021 gtgcagctca cctccagcac actgaccaac cctaggcaga gccccgtgga ggcccaggac
     1081 cgggagacct ggggcaagaa aatcgacttc ctgctcagcg tgatcggctt cgccgtcgac
     1141 ctggctaacg tttggcggtt cccctacctg tgctacaaga acggcggagg cgctttcctg
     1201 gtgccctacc tcctctttat ggtgatcgct ggcatgcccc tgttctacat ggagctggcc
     1261 ctcggccagt ttaaccggga gggagccgca ggtgtgtgga agatctgccc catcctcaaa
     1321 ggcgtgggat tcaccgtgat cctgatcagc ctgtacgtag gcttctttta caacgtcatc
     1381 attgcctggg ctctccacta cctgttctcg agcttcacca cagagctgcc ctggatccac
     1441 tgcaacaata gctggaacag ccccaactgc agcgatgcgc accccggcga cagctccgga
     1501 gacagctcag gcctgaacga caccttcgga accacacccg ccgctgaata cttcgaacgg
     1561 ggcgtgctgc acctccatca gagccacggc atcgatgacc ttggtcctcc caggtggcag
     1621 ctgaccgcct gtctggtgct cgtcatcgtg ctgctctact tcagcctgtg gaagggcgtg
     1681 aagacaagcg gcaaggtggt ctggatcacc gccacaatgc catatgtggt cctgaccgcc
     1741 ctgctcctgc ggggggtgac cctgcctggg gccatcgacg gcattcgggc ctacctgagc
     1801 gtggacttct accggctgtg cgaggctagc gtgtggatcg acgctgccac tcaagtgtgc
     1861 ttcagcctgg gcgtgggatt cggggtcctg atcgccttca gctcctacaa caagttcacc
     1921 aacaattgct accgcgacgc catcgtcacc acaagcatca actccctgac tagtttttcg
     1981 agcggctttg tcgtgttcag ctttctggga tacatggccc agaagcacag cgtgcccatc
     2041 ggtgacgtcg ccaaggacgg ccccgggctg atcttcatta tctaccctga ggccatcgct
     2101 accctgcccc tgagctccgc ctgggctgtc gtgttcttta tcatgctgct caccctcggc
     2161 atcgacagcg ctatgggagg catggagagc gtcatcaccg gcctgatcga cgaattccag
     2221 ctcctgcacc ggcatcgcga gctgttcacc ctgttcatcg tgctggccac cttccttctg
     2281 agcctgttct gcgtgaccaa cggcggaatc tacgtgttca ccctcctgga ccacttcgct
     2341 gccggcacct caatattgtt cggcgtgctg atcgaggcga tcggcgtggc ctggttctac
     2401 ggagtgggcc agttcagcga cgatatccag cagatgaccg gtcagcggcc cagcctgtac
     2461 tggcggctgt gctggaagct cgtctccccc tgcttcctcc tgttcgtggt cgttgtgtca
     2521 attgtgacct tccggcctcc ccactacggc gcctacatct tccccgactg ggccaatgca
     2581 ttgggctggg tgatcgcaac cagctccatg gctatggtgc ccatctacgc tgcctacaag
     2641 ttctgtagcc tgcccggaag cttccgggag aagctggcgt acgctatcgc ccccgagaag
     2701 gaccgggagc tggtggaccg cggcgaggtg cggcagttca ccctgcggca ctggctgaag
     2761 gtgtgaagag agtctagaag cggccgcgtc gacggaattc agtggatcca ctagtaacgg
     2821 ccgccagtgt gctggaatta attcgctgtc tgcgagggcc agctgttggg gtgagtactc
     2881 cctctcaaaa gcgggcatga cttctgcgct aagattgtca gtttccaaaa acgaggagga
     2941 tttgatattc acctggcccg cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga
     3001 aaagacaatc tttttgttgt caagcttgag gtgtggcagg cttgagatct ggccatacac
     3061 ttgagtgaca atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     3121 ctgcaggtcg agcatgcatc tagggcggcc ggccgcacta gaggaattcc gcccctctcc
     3181 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgtgtttgt
     3241 ctatatgtga ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     3301 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     3361 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     3421 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     3481 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3541 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtagtcaaca aggggctgaa
     3601 ggatgcccag aaggtacccc attgtatggg aatctgatct ggggcctcgg tgcacatgct
     3661 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3721 ttttcctttg aaaaacacga tgataagctt gccacaaccc gtaccaaaga tggatagatc
     3781 cggaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3841 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3901 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3961 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     4021 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4081 agacctgcct gaaaccgaac tgcccgctgt tctgcaccgg tcgcggaggc catggatgcg
     4141 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     4201 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     4261 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     4321 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     4381 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     4441 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     4501 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg
     4561 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     4621 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     4681 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     4741 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     4801 tagagtagat gccgaccgaa caagagctga tttcgagaac gcctcagcca gcaactcgcg
     4861 cgagcctagc aaggcaaatg cgagagaacg gccttacgct tggtggcaca gttctcgtcc
     4921 acagttcgct aagctcgctc ggctgggtcg cgggagggcc ggtcgcagtg attcaggccc
     4981 ttctggattg tgttggtccc cagggcacga ttgtcatgcc cacgcactcg ggtgatctga
     5041 ctgatcccgc agattggaga tcgccgcccg tgcctgccga ttgggtgcag atctagagct
     5101 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5161 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5221 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5281 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     5341 gcttctgagg cggaaagaac cagctggggc tcgagtgcat tctagttgtg gtttgtccaa
     5401 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5521 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6061 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6301 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6541 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6601 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6661 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6721 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6781 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6841 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6901 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6961 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7021 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7081 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7141 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7201 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7261 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7321 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7381 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7441 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7501 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7561 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7621 cgtc
//
