LOCUS       sequence-445138-        7229 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 7229)
  TITLE     Susan Amara Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7229)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1-hDAT
FEATURES             Location/Qualifiers
     source          1..7229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        116..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     650..670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     744..763
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        744..762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1239..1250
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     primer_bind     complement(2701..2718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2707..2931
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2977..3405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3064..3083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3274..3295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3414..3434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3419..3748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3599..3734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3661..3680
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3815..4609
                     /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(3869..3888)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4479..4498
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4783..4904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4820..4839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4874..4893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4953..4969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4953..4969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4966..4988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4977..4993
                     /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(5001..5007,5008..5025,5026..5031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5046..5067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5184..5201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5355..5943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5435..5454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6114..6905,6906..6974))
                     /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     6737..6756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6975..7079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7154..7173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
       61 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      721 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      781 cgtttaaact taagcttggt accaccatga gtaagagcaa atgctccgtg ggactcatgt
      841 cttccgtggt ggccccggct aaggagccca atgccgtggg cccgaaggag gtggagctca
      901 tccttgtcaa ggagcagaac ggagtgcagc tcaccagctc caccctcacc aacccgcggc
      961 agagccccgt ggaggcccag gatcgggaga cctggggcaa gaagatcgac tttctcctgt
     1021 ccgtcattgg ctttgctgtg gacctggcca acgtctggcg gttcccctac ctgtgctaca
     1081 aaaatggtgg cggtgccttc ctggtcccct acctgctctt catggtcatt gctgggatgc
     1141 cacttttcta catggagctg gccctcggcc agttcaacag ggaaggggcc gctggtgtct
     1201 ggaagatctg ccccatactg aaaggtgtgg gcttcacggt catcctcatc tcactgtatg
     1261 tcggcttctt ctacaacgtc atcatcgcct gggcgctgca ctatctcttc tcctccttca
     1321 ccacggagct cccctggatc cactgcaaca actcctggaa cagccccaac tgctcggatg
     1381 cccatcctgg tgactccagt ggagacagct cgggcctcaa cgacactttt gggaccacac
     1441 ctgctgccga gtactttgaa cgtggcgtgc tgcacctcca ccagagccat ggcatcgacg
     1501 acctggggcc tccgcggtgg cagctcacag cctgcctggt gctggtcatc gtgctgctct
     1561 acttcagcct ctggaagggc gtgaagacct cagggaaggt ggtatggatc acagccacca
     1621 tgccatacgt ggtcctcact gccctgctcc tgcgtggggt caccctccct ggagccatag
     1681 acggcatcag agcatacctg agcgttgact tctaccggct ctgcgaggcg tctgtttgga
     1741 ttgacgcggc cacccaggtg tgcttctccc tgggcgtggg gttcggggtg ctgatcgcct
     1801 tctccagcta caacaagttc accaacaact gctacaggga cgcgattgtc accacctcca
     1861 tcaactccct gacgagcttc tcctccggct tcgtcgtctt ctccttcctg gggtacatgg
     1921 cacagaagca cagtgtgccc atcggggacg tggccaagga cgggccaggg ctgatcttca
     1981 tcatctaccc ggaagccatc gccacgctcc ctctgtcctc agcctgggcc gtggtcttct
     2041 tcatcatgct gctcaccctg ggtatcgaca gcgccatggg tggtatggag tcagtgatca
     2101 ccgggctcat cgatgagttc cagctgctgc acagacaccg tgagctcttc acgctcttca
     2161 tcgtcctggc gaccttcctc ctgtccctgt tctgcgtcac caacggtggc atctacgtct
     2221 tcacgctcct ggaccatttt gcagccggca cgtccatcct ctttggagtg ctcatcgaag
     2281 ccatcggagt ggcctggttc tatggtgttg ggcagttcag cgacgacatc cagcagatga
     2341 ccgggcagcg gcccagcctg tactggcggc tgtgctggaa gctggtcagc ccctgctttc
     2401 tcctgttcgt ggtcgtggtc agcattgtga ccttcagacc cccccactac ggagcctaca
     2461 tcttccccga ctgggccaac gcgctgggct gggtcatcgc cacatcctcc atggccatgg
     2521 tgcccatcta tgcggcctac aagttctgca gcctgcctgg gtcctttcga gagaaactgg
     2581 cctacgccat tgcacccgag aaggaccgtg agctggtgga cagaggggag gtgcgccagt
     2641 tcacgctccg ccactggctc aaggtgtagt ctagagggcc cgtttaaacc cgctgatcag
     2701 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2761 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2821 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2881 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2941 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3001 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3061 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3181 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3301 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3421 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3481 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3601 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3661 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3721 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3781 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3841 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3901 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3961 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4021 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4081 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4141 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4201 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4261 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4321 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4381 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4441 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4501 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4561 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4621 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4681 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4741 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4801 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4861 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4921 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4981 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5041 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7081 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7141 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7201 cttgtgtgtt ggaggtcgct gagtagtgc
//
