LOCUS       Exported                8025 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    hSERT pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8025)
  AUTHORS   Ramamoorthy S, Bauman AL, Moore KR, Han H, Yang-Feng T, Chang AS, 
            Ganapathy V, Blakely RD
  TITLE     Antidepressant- and cocaine-sensitive human serotonin transporter: 
            molecular cloning, expression, and chromosomal localization.
  JOURNAL   Proc Natl Acad Sci U S A. 1993 Mar 15. 90(6):2542-6.
  PUBMED    7681602
REFERENCE   2  (bases 1 to 8025)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8025
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     primer_bind     972..988
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     972..988
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3538..3554)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3578..3596)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3579..3596)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3616..3633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3622..3846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3892..4320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3979..3998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4189..4210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4329..4349)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4334..4663
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4514..4649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4576..4595
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4730..5524
                     /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(4784..4803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5394..5413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5698..5819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5735..5754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5789..5808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5868..5884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5868..5884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5881..5903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5892..5908
                     /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(5916..5946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5961..5982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6099..6116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6270..6858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6350..6369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7029..7889)
                     /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     7652..7671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7890..7994)
                     /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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
      961 ctggcggccg ctctagaact agtggatccc ccgggctgca ggaattccca aatccaagca
     1021 cccagagatc aattgggatc cttggcagat ggacatcagt gtcatttact aaccagcagg
     1081 atggagacga cgcccttgaa ttctcagaag cagctatcag cgtgtgaaga tggagaagat
     1141 tgtcaggaaa acggagttct acagaaggtt gttcccaccc caggggacaa agtggagtcc
     1201 gggcaaatat ccaatgggta ctcagcagtt ccaagtcctg gtgcgggaga tgacacacgg
     1261 cactctatcc cagcgaccac caccacccta gtggctgagc ttcatcaagg ggaacgggag
     1321 acctggggca agaaggtgga tttccttctc tcagtgattg gctatgctgt ggacctgggc
     1381 aatgtctggc gcttccccta catatgttac cagaatggag ggggggcatt cctcctcccc
     1441 tacaccatca tggccatttt tgggggaatc ccgctctttt acatggagct cgcactggga
     1501 cagtaccacc gaaatggatg catttcaata tggaggaaaa tctgcccgat tttcaaaggg
     1561 attggttatg ccatctgcat cattgccttt tacattgctt cctactacaa caccatcatg
     1621 gcctgggcgc tatactacct catctcctcc ttcacggacc agctgccctg gaccagctgc
     1681 aagaactcct ggaacactgg caactgcacc aattacttct ccgaggacaa catcacctgg
     1741 accctccatt ccacgtcccc tgctgaagaa ttttacacgc gccacgtcct gcagatccac
     1801 cggtctaagg ggctccagga cctggggggc atcagctggc agctggccct ctgcatcatg
     1861 ctgatcttca ctgttatcta cttcagcatc tggaaaggcg tcaagacctc tggcaaggtg
     1921 gtgtgggtga cagccacctt cccttatatc atcctttctg tcctgctggt gaggggtgcc
     1981 accctccctg gagcctggag gggtgttctc ttctacttga aacccaattg gcagaaactc
     2041 ctggagacag gggtgtggat agatgcagcc gctcagatct tcttctctct tggtccgggc
     2101 tttggggtcc tgctggcttt tgctagctac aacaagttca acaacaactg ctaccaagat
     2161 gccctggtga ccagcgtggt gaactgcatg acgagcttcg tttcgggatt tgtcatcttc
     2221 acagtgctcg gttacatggc tgagatgagg aatgaagatg tgtctgaggt ggccaaagac
     2281 gcaggtccca gcctcctctt catcacgtat gcagaagcga tagccaacat gccagcgtcc
     2341 actttctttg ccatcatctt ctttctgatg ttaatcacgc tgggcttgga cagcacgttt
     2401 gcaggcttgg agggggtgat cacggctgtg ctggatgagt tcccacacgt ctgggccaag
     2461 cgccgggagc ggttcgtgct cgccgtggtc atcacctgct tctttggatc cctggtcacc
     2521 ctgacttttg gaggggccta cgtggtgaag ctgctggagg agtatgccac ggggcccgca
     2581 gtgctcactg tcgcgctgat cgaagcagtc gctgtgtctt ggttctatgg catcactcag
     2641 ttctgcaggg acgtgaagga aatgctcggc ttcagcccgg ggtggttctg gaggatctgc
     2701 tgggtggcca tcagccctct gtttctcctg ttcatcattt gcagttttct gatgagcccg
     2761 ccacaactac gacttttcca atataattat ccttactgga gtatcatctt gggttactgc
     2821 ataggaacct catctttcat ttgcatcccc acatatatag cttatcggtt gatcatcact
     2881 ccagggacat ttaaagagcg tattattaaa agtattaccc cagaaacacc aacagaaatt
     2941 ccttgtgggg acatccgctt gaatgctgtg taacacactc accgagagga aaaaggcttc
     3001 tccacaacct cctcctccag ttctgatgag gcacgcctgc cttctcccct ccaagtgaat
     3061 gagtttccag ctaagcctga tgatggaagg gccttctcca cagggacaca gtctggtgcc
     3121 cagactcaag gcctccagcc acttatttcc atggattccc ctggacatat tcccatggta
     3181 gactgtgaca cagctgagct ggcctatttt ggacgtgtga ggatgtggat ggaggtgatg
     3241 aaaaccaccc tatcatcagt taggattagg tttagaatca agtctgtgaa agtctcctgt
     3301 atcatttctt ggtatgatca ttggtatctg atatctgttt gcttctaaag gtttcactgt
     3361 tcatgaatac gtaaactgcg taggagagaa cagggatgct atctcgctag ccatatattt
     3421 tctgagtagc atatagaatt ttattgctgg aatctactag aaccttctaa tccatgtgct
     3481 gctgtggcat caggaaagga agatgtaaga agctaaggaa ttcgatatca agcttatcga
     3541 taccgtcgac ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta
     3601 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3661 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3721 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3781 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3841 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3901 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3961 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4021 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4081 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4141 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4201 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4261 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4321 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4441 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4501 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4561 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4621 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4681 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4741 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4801 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4861 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4921 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4981 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5041 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5101 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5161 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5221 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5281 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5341 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5401 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5461 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5521 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5581 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5641 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7021 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7081 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7141 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7201 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7261 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7321 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7381 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7441 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7501 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7561 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7621 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7681 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7741 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7801 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7861 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7921 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7981 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
